Production and Characterization of Monoclonal and Polyclonal Antibodies Against Truncated Recombinant Dickkopf-1 as a Candidate Biomarker.
Authors: Malaei F, Rasaee MJ, Paknejad M, Latifi AM, Rahbarizadeh F Abstract Several studies have reported an increased serum level of Dickkopf (DKK-1) protein in a variety of cancers, including multiple myeloma, lung, colorectal, bone loss, and Alzheimer's disease. This protein has potential to be used as a biomarker for the diagnosis of some cancers, especially bone loss in multiple myeloma. In the present study, to measure the concentration level of DKK-1 protein, rabbit polyclonal antibody (pAb) and mouse monoclonal antibodies (mAbs) were produced against this protein. New Zealand white rabbits and BALB/c mice ...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 30, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Crude Inactivated Influenza A Virus Adjuvated with a Bispecific Antibody Complex Targeting Chicken CD40 and AIV M2e Confers Protection Against Lethal HPAI Challenge in Chickens.
Authors: Vuong CN, Chou WK, Briggs W, Faulkner O, Wolfenden A, Jonas M, Kapczynski DR, Hargis BM, Bielke LR, Berghman LR Abstract In vivo targeting an immunogen to the CD40 receptor expressed on professional antigen-presenting cells (APCs) dramatically enhances speed, magnitude, and quality of the immune response. Our previous evaluation of this strategy in poultry was limited to immunogenicity studies using CD40-targeted synthetic peptides, which demonstrated significant antigen-specific serum IgG and tracheal IgA levels
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 30, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Galactose-Deficient IgA1-Specific Antibody Recognizes GalNAc-Modified Unique Epitope on Hinge Region of IgA1.
In this study, we aimed to analyze the epitope of KM55 mAb using synthesized peptides corresponding to the hinge region of IgA1 with GalNAc moiety on putative glycosylated Ser/Thr residues, which are Thr225, Thr228, Ser230, Ser232, and Thr236. Binding analysis to single GalNAc-modified hinge region peptide of IgA1 showed that Thr225 with GalNAc is required for recognition of KM55. PST(GalNAC)PP motif was required for KM55 mAb to recognize hinge region peptide of IgA1 which is shown by binding assay with deletion peptide. This result was confirmed by binding of KM55 mAb against peptide with GalNAc at Thr233, which resulted ...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 21, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Immunohistochemical Detection of Sheep Podoplanin Using an Antibovine Podoplanin Monoclonal Antibody PMab-44.
In this study, we investigated possible cross-reactions between anti-PDPN mAbs and sheep PDPN. Type I alveolar cells from sheep lung were strongly detected by PMab-44 using immunohistochemical analyses. These results indicate that PMab-44 may be useful for the detection of sheep PDPN. PMID: 30570359 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 21, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Detection of Alpaca Podoplanin by Immunohistochemistry Using the Antibovine Podoplanin Monoclonal Antibody PMab-44.
In this study, we investigated the potential cross-reaction between established antibovine PDPN mAbs and alpaca PDPN. Using immunohistochemical analysis, type I alveolar cells of the alpaca lungs were detected by the antibovine PDPN mAb, PMab-44. These results indicate that PMab-44 may be useful for the detection of alpaca PDPN. PMID: 30570440 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 21, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Association of CDH1 and TERT Single-Nucleotide Polymorphisms with Susceptibility to Familial Breast Cancer Risk.
This study aimed to analyze the association of CDH1 and TERT single-nucleotide polymorphisms with susceptibility to familial BC (FBC) risk in the Iranian patients. One hundred five patients with FBC and 110 non-FBC (NFBC) were genotyped to elucidate the potential association between CDH1 rs5030625 polymorphism and TERT rs2736098 polymorphism by polymerase chain reaction-restriction fragment length polymorphism. Then, results were evaluated by electrophoresis and Epi Info(™) 2012 software. A significant association was found between CDH1 rs5030625 GAGA genotype and FBC risk. Compared with the control group, the FBC pa...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 15, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Anti-Bovine Podoplanin Monoclonal Antibody PMab-44 Detects Goat Podoplanin in Immunohistochemistry.
In this study, we investigated whether these anti-PDPN mAbs cross-react with goat PDPN (gPDPN). Flow cytometry demonstrated that the anti-bovine PDPN mAb PMab-44 (IgG1, kappa) reacts with gPDPN, which is overexpressed in CHO-K1 cells. Using immunohistochemical analysis, type I alveolar cells of goat lung were strongly detected by PMab-44. These results indicate that PMab-44 is useful for investigating gPDPN. PMID: 30457925 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - November 21, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin.
In this study, we developed and produced several mouse-canine chimeric antibodies originating from PMab-38. A mouse-canine chimeric antibody of subclass A (P38A) and a mouse-canine chimeric antibody of subclass B (P38B) were transiently produced using ExpiCHO-S cells. Core-fucose-deficient P38B (P38Bf) was developed using FUT8 knockout ExpiCHO-S cells. We compared the binding affinities, antibody-dependent cellular cytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC) of P38A, P38B, and P38Bf against Chinese hamster ovary (CHO)/dPDPN cells. Flow cytometry analysis showed that the KD of P38A, P38B, and P38Bf were...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - October 28, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Novel Monoclonal Antibodies Against Native Human Collectin L1, Using the Heteromeric Complex CL-LK as Immunogen.
Authors: Aagaard JB, Bjerrum KB, Graversen JH, Henriksen ML, Hansen SWK Abstract Collectin LK (CL-LK) is a recently described collectin complex, which upon binding to microbial glycoconjugates, activates the lectin pathway of the complement system, and thereby contributes to the removal of invading microorganisms. The complex consists of the two collectins; Collectin K1 (kidney 1 alias CL-11) and Collectin L1 (liver 1 alias CL-10). At present, most efforts have been made on the characterization of CL-K1, and little is known about the function of CL-L1 and its association with diseases. Deficiency of either of the t...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - October 28, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Detection of Tiger Podoplanin Using the Anti-Cat Podoplanin Monoclonal Antibody PMab-52.
In this study, we investigated the possible crossreaction between these anti-PDPN mAbs and tiger PDPN. Flow cytometry and western blot analyses revealed that the anti-cat PDPN mAb PMab-52 (IgM, kappa) reacted with tiger PDPN, which is overexpressed in Chinese hamster ovary-K1 cells. Using immunohistochemical analysis, type I alveolar cells of the tiger lung were strongly detected by PMab-52. These results indicate that PMab-52 may be useful for the detection of tiger PDPN. PMID: 30362928 [PubMed - in process] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - October 28, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generation of a Monoclonal Antibody Against Staphylococcal Superantigen-Like Protein 5 (SSL5) That Discriminates SSL5 from Other SSL Proteins.
In this study, we established a mouse hybridoma clone (30G5C) producing a monoclonal antibody specific for SSL5. In immunoblotting analysis using recombinant His-tagged SSL1 to SSL14 (His-SSLs), the antibody was found to specifically recognize SSL5 without crossreactivity with other His-SSLs. The antibody bound to the C-terminal region of SSL5 (β-grasp domain), but did not interfere with the binding of SSL5 to MMP-9, suggesting that this antibody is useful for identification of SSL5-producing S. aureus and screening for inhibitors of the SSL5/MMP-9 complex formation. PMID: 30362929 [PubMed - in process] (Source: M...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - October 28, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

The Augmenting Effects of the tDNA Insulator on Stable Expression of Monoclonal Antibody in Chinese Hamster Ovary Cells.
Authors: Naderi F, Hashemi M, Bayat H, Mohammadian O, Pourmaleki E, Etemadzadeh MH, Rahimpour A Abstract Cell line development is one of the most critical steps in the production of complex recombinant therapeutic proteins such as monoclonal antibodies in mammalian cells. Generation of industrial cell lines is mainly based on the time-consuming and laborious process of selection and screening of a large number of clones. With the increasing demand for therapeutic proteins during the past years, more effort is invested to improve the efficiency of cell line development. In line with this premise, several studies emp...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - October 28, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

DgMab-6: Antihuman DGK γ Monoclonal Antibody for Immunocytochemistry.
In this study, we developed a novel anti-DGKγ monoclonal antibody, DgMab-6, which is very useful in immunocytochemistry of human cultured cells. PMID: 30362931 [PubMed - in process] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - October 28, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Establishment of Monoclonal Antibody PMab-202 Against Horse Podoplanin.
In this study, we immunized mice with synthetic horse PDPN peptides and developed anti-horse PDPN mAbs. One of the established mAbs, PMab-202 (IgG1, kappa), was specifically able to detect horse PDPN in Chinese hamster ovary/horse PDPN (CHO/horPDPN) cells in flow cytometry experiments. PMab-202 was also able to detect endogenous horse PDPN expressed in and a horse kidney cell line, FHK-Tcl3.1, in flow cytometry and Western blot analyses. PMab-202 is expected to prove useful in investigating the function of horse PDPN. PMID: 30362932 [PubMed - in process] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - October 28, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Advancing Freund's and AddaVax Adjuvant Regimens Using CpG Oligodeoxynucleotides.
Authors: Hawksworth D Abstract Adjuvant can play an important role in vaccine formulation by aiding in the development of a robust immune response. In our hybridoma development work, we typically use both Freund's and AddaVax™ adjuvant regimens for mouse immunizations. While we have repeatedly shown success with our protocols, we continually seek to improve upon the titer and affinity of the serum antibody response. To that end, we evaluated the use of CpG oligodeoxynucleotides (CpG-DNA), a B cell stimulant, in our adjuvant regimens. Mice were immunized using our standard Freund's protocol (Adjulite Complete ...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - October 5, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal Antibody Specific for Ovarian Tumor Domain-Containing Protein 1.
Authors: PMID: 30161019 [PubMed - in process] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - September 1, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generation of Rat Monoclonal Antibodies Against a Deubiquitinase, Ovarian Tumor Domain-Containing Protein 1.
Authors: Oikawa D, Shiota M, Goto E, Komakura K, Wanibuchi H, Tokunaga F Abstract Ovarian tumor domain-containing protein 1 (OTUD1), an OTU-family deubiquitinating enzyme, has been reported to be involved in cancer progression through the regulation of p53 and SMAD7. However, the precise pathophysiological functions of OTUD1 remain elusive. Here, we report the establishment of OTUD1-specific monoclonal antibodies (mAbs), using the rat medial iliac lymph node method. The generated antibodies recognize the N-terminal portion (aa. 1-290) of human and mouse OTUD1 proteins. In addition, immunofluorescent staining and su...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - August 23, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development and Characterization of Monoclonal Antibodies Against Nitro-166Tyrosine of High-Density Lipoprotein: Apolipoprotein A1.
In this study, we developed monoclonal antibodies (mAbs) specific to modified ApoA1 with its tyrosine residue at the 166th position nitrated to 3-nitrotyrosine. A 20 amino acid peptide around the modification of interest was designed using an antigenicity prediction tool. The peptide was custom synthesized with ovalbumin as conjugate and used as an antigen to immunize BALB/c mice. Hybridomas were obtained by fusion of Sp2/0 mouse myeloma cells with spleen cells from the immunized mouse. A hybridoma clone 2E5B7, thus developed and characterized, was found to secrete mAb of the desired specificity and sensitivity against nit...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - August 23, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Evidence for Multiple Applications of Monoclonal Antibody 5G10.
In this study, we explored multiple functions of 5G10. A short peptide QRVRELAV (designated T5) derived from SF in either terminal of proteins was specifically recognized by 5G10. T5 tag expressed in eukaryotic cell was also detected by 5G10 when analyzed by Western blot, immunofluorescence assay (IFA), and fluorescent-activated cell sorting (FACS). The result of the co-immunoprecipitation assay showed that 5G10 as a bait antibody dragged out the complex of enterovirus 71 (EV71) 2A and mitochondrial antiviral signaling (MAVS) protein. More importantly, 5G10 helped to purify fusion proteins T5-tagged (EV71) 2A and T5-Japane...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - August 23, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Elucidation of Critical Epitope of Anti-Rat Podoplanin Monoclonal Antibody PMab-2.
Authors: Furusawa Y, Yamada S, Itai S, Nakamura T, Fukui M, Harada H, Kaneko MK, Kato Y Abstract Rat podoplanin (rPDPN) is a recognized lymphatic endothelial cell marker and is expressed on the podocytes of kidney and type I lung alveolar cells. rPDPN is a type I transmembrane sialoglycoprotein and induces platelet aggregation via the C-type lectin-like receptor-2 of platelets. It comprises four platelet aggregation-stimulating (PLAG) domains: PLAG1-3, present in the N-terminus, and PLAG4, in the center of the PDPN protein. Previously, we developed a mouse anti-rPDPN monoclonal antibody clone, PMab-2, by immunizing...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - August 11, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Epitope Mapping of Anti-Telomerase Reverse Transcriptase Monoclonal Antibodies.
Authors: Furusawa Y, Itai S, Yamada S, Kaneko MK, Kato Y Abstract Telomerase reverse transcriptase (TERT) is the main catalytic subunit of telomerase-associated protein machinery. Upregulation of TERT at the transcriptional level results in immortal cell phenotype associated with cancer. To date we have developed eight anti-TERT monoclonal antibodies (mAbs) (TMab-4, TMab-5, TMab-6, TMab-7, TMab-9, TMab-10, TMab-11, and TMab-12) by immunizing mice with synthetic peptides (302-QHHAGPPSTSRPPRPWDTPC-321) of TERT. We further characterized those epitopes using enzyme-linked immunosorbent assay, and here we discuss the cr...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - July 14, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal Antibody MAb SZ-189 Against Recombinant Human CLEC-2 Protein.
Authors: PMID: 29939835 [PubMed - in process] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - June 27, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Thinking Cancer.
Authors: Pashov A, Hernandez Puente CV, Ibrahim SM, Monzavi-Karbassi B, Makhoul I, Kieber-Emmons T Abstract Evolutionary theories are necessarily invoked for understanding cancer development at the level of species, at the level of cells and tissues, and for developing effective therapies. It is crucial to view cancer in a Darwinian light, where the differential survival of individual cells is based on heritable variations. In the process of this somatic evolution, multicellularity controls are overridden by cancer cells, which become increasingly autonomous. Ecological epigenetics also helps understand how rogue c...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - June 27, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Non-Invasive Immunofluorescence Assessment of Lycopene Supplementation Status in Skin Smears.
Authors: Petyaev IM, Zigangirova NA, Pristensky D, Chernyshova M, Tsibezov VV, Chalyk NE, Morgunova EY, Kyle NH, Bashmakov YK Abstract Circulating lycopene level is negatively associated with the prevalence of cardiovascular disease, cancers (prostate and breast), type 2 diabetes mellitus, and aging. Traditionally, lycopene is measured in biological specimens by a combination of high-performance liquid chromatography (HPLC) and mass spectrometry methods. Moreover, as we recently reported, tissue/cell lycopene depositions can be observed by the immunohistochemistry method with a newly developed monoclonal antibody (...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - June 16, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Association with Monoclonal Antibody Promotes Intracellular Delivery of Lycopene.
Authors: Petyaev IM, Zigangirova NA, Tsibezov VV, Morgunova EY, Bondareva NE, Kyle NH, Bashmakov YK Abstract Incubation of B10.MLM cells, a cell line of alveolar macrophages, with lycopene, a carotenoid, leads to an increase of lycopene content in their microsomal fraction. That increase was higher and developed faster when the cells were incubated with immune complexes formed by lycopene and mAb 6B9 (L-6B9 mAb), a monoclonal hapten-specific antibody raised against lycopene, as compared with dimethyl sulfoxide (DMSO)-dissolved lycopene (DMSO-L). Moreover, incubation of B10.MLM cells with L-6B9 mAb complexes was acc...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - June 16, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

A Bioassay for Optimization of Macrophage-Conditioned Medium as a Culture Supplement to Promote Hybridoma Cell Survival and Growth.
In conclusion, the RMH359 cell bioassay provides a specific and sensitive assessment of MCM bioactivity in support of hybridoma cell survival and growth. PMID: 29901420 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - June 16, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generation of Rat Monoclonal Antibodies Specific for DZIP3.
In this study, we report the establishment of DZIP3-specific monoclonal antibodies (MAbs), using the rat medial iliac lymph node method. In immunoblotting analyses, our antibodies detected endogenous human and mouse DZIP3. In addition, immunoprecipitation analyses revealed the availability of these antibodies for human or mouse DZIP3. Thus, these MAbs will be available to elucidate cellular functions of DZIP3. PMID: 29812999 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - May 31, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Preparation and Application of a Monoclonal Antibody Against Chicken TRIM62.
Authors: Wang X, Zhao Y, Li L, Cheng Z, Wang G Abstract TRIM62 is a member of the tripartite interaction motif (TRIM) family and exerts crucial roles in innate immune response and cancer. To investigate the relationship between its distribution and avian retrovirus replication, in the present study, a monoclonal antibody (mAb) against chicken TRIM62 was generated. The open reading frame of chicken TRIM62 was amplified by polymerase chain reaction (PCR) and inserted into the expression vector pET-28a. The recombinant expression vectors were transformed into Escherichia coli BL21 (DE3). Then recombinant protein His-T...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - May 31, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Elucidation of the TMab-6 Monoclonal Antibody Epitope Against Telomerase Reverse Transcriptase.
In this study, enzyme-linked immunosorbent assay and immunohistochemistry were utilized for investigating the epitope of TMab-6. The findings revealed that the critical epitope of TMab-6 is the TERT sequence PSTSRPPRPWD; Thr310 and Ser311 of TERT are especially significant amino acids for TMab-6 recognition. PMID: 29723111 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - May 4, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

The Characterization of Monoclonal Antibodies to Mouse TLT-1 Suggests That TLT-1 Plays a Role in Wound Healing.
Authors: Manfredi B, Morales-Ortíz J, Díaz-Díaz LM, Hernandez-Matias L, Barreto-Vázquez D, Menéndez-Pérez J, Rodríguez-Cordero JA, Villalobos-Santos JC, Santiago-Rivera E, Rivera-Dompenciel A, Lozada-Delgado EL, Kuchibhotla M, Carrasquillo-Carrión K, Roche-Lima A, Washington AV Abstract Platelets play a vital role in hemostasis and inflammation. The membrane receptor TREM-like transcript-1 (TLT-1) is involved in platelet aggregation, bleeding, and inflammation, and it is localized in the α-granules of platelets. Upon platelet activation, TLT-1 is release...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - May 1, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development of Monoclonal Antibodies Specifically Recognizing the Nonstructural Protein 12 of Type 2 Porcine Reproductive and Respiratory Syndrome Virus.
In this study, we generated and identified the monoclonal antibody (mAb) against PRRSV Nsp12. Six strains of hybridoma cells named 2B10, 2B12, 5E1, 5G6, 5E7, and 8B2 secreting anti-Nsp12 mAbs were obtained by the hybridoma technique. All the mAbs were specifically reacted with PRRSV by indirect immunofluorescence assay and four of them (2B12, 5E1, 5G6, and 5E7) were specifically reacted by Western blot. Furthermore, the 5E7 specifically recognized multiple type 2 PRRSV strains, including highly pathogenic and classical PRRSV strains, but not type 1 PRRSV strain. Taken together, the mAbs against Nsp12 provide a valuable too...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - May 1, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies.
We report an immunization technique that can update the production of monoclonal antibodies (mAbs): the multiple tolerization subtractive immunization (MTSI). A total of 10 BALB/C mice were used. Animals in group 1 received one inoculation of RWPE-1 cells (nontumoral), followed by cyclophosphamide, and then received serial inoculations of nonirradiated PC3 cells (tumoral). Animals in group 2 received our MTSI protocol, as follows: one inoculation of RWPE-1 cells, followed by cyclophosphamide (Cy). This whole tolerization step was repeated three other times, with 14-day intervals between the last Cy exposure and the next RW...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - May 1, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Quantification of Histidine-Rich Protein 3 of Plasmodium falciparum.
Authors: Palani B Abstract Malaria is a life-threatening infectious disease and continues to be a major public health crisis in many parts of the tropical world. Plasmodium falciparum is responsible for the majority of mortality and morbidity associated with malaria. During the intraerythrocytic cycle, P. falciparum releases three proteins with high histidine content as follows: histidine-rich protein 1 (HRP1), histidine-rich protein 2 (HRP2), and histidine-rich protein 3 (HRP3). Currently, most of the diagnostic tests of P. falciparum infection target HRP2, and a number of monoclonal antibodies (mAbs) against HRP2...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - April 14, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

TX99 Is a Neutralizing Monoclonal Antibody Against Mouse TIGIT.
In this study, we generated five clones of anti-mTIGIT mAbs, designated TX99, TX100, TX103, TX104, and TX105. We show that TX99 and TX100 showed the strongest binding to TIGIT. We also show that TX99 interfered with the interaction between TIGIT and CD155 and increased NK cell-mediated cytotoxicity against CD155-expressing RMA-S cells. Thus, TX99 is a unique neutralizing mAb that can be used for studies of mTIGIT functions. PMID: 29648914 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - April 14, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Anti-Podocalyxin Monoclonal Antibody 47-mG2a Detects Lung Cancers by Immunohistochemistry.
In this study, we investigated the utility of PcMab-47 and 47-mG2a for the immunohistochemical analyses of lung cancers. PcMab-47 stained 51/70 (72.9%) cases of lung cancer, whereas 47-mG2a stained 59/70 (84.3%) cases, indicating that the latter antibody is more sensitive and is useful for detecting PODXL in lung cancers. PMID: 29649375 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - April 14, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

47-mG2a: A Mouse IgG2a-Type of PcMab-47 Useful for Detecting Podocalyxin in Esophageal Cancers by Immunohistochemistry.
Authors: Kaneko MK, Itai S, Yamada S, Kato Y Abstract Esophageal cancer is one of the highly malignant cancers. It comprises two of the most common histological tumor types: squamous cell carcinoma (SCC) and adenocarcinoma. SCC accounts for about 90% of esophageal cancers. Despite developments in treatment strategies, the prognosis and survival rate remain poor. Podocalyxin (PODXL) is a highly glycosylated type-I transmembrane protein. It is expressed in normal tissues such as kidney, heart, breast, and pancreas. Upregulation of PODXL correlates with tumor progression, invasion, and metastasis. Therefore, this glyc...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - April 11, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal Antibody Against HA Protein of the European Avian-Like H1N1 Swine Influenza Virus.
Authors: Wang Q, Wang SY, Zhang P, Liu XM, Yu LX, Shan TL, Tong W, Zhou YJ, Li GX, Zheng H, Gao F, Jiang YF, Kong N, Li LW, Tong GZ, Yu H Abstract The purified whole-virus proteins derived from A/swine/Shanghai/1/2014 (H1N1) (SH1) were chosen to immunize BALB/c mice to prepare the monoclonal antibody (MAb) against hemagglutinin (HA) protein of an European avian-like (EA) H1N1 swine influenza virus (SIV). After cloning three times by limiting dilution, one strain of hybridoma cells named 3C7 secreting anti-HA protein MAb was obtained by hybridoma technique. The results of indirect immunofluorescence assay and wester...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - April 11, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Epitope Mapping of Monoclonal Antibody PMab-48 Against Dog Podoplanin.
In this study, an enzyme-linked immunosorbent assay and flow cytometry were used to investigate epitopes of PMab-48. The results revealed that the critical epitope of PMab-48 comprises Asp29, Asp30, Ile31, Ile32, and Pro33 of dPDPN. PMID: 29608407 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - April 4, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal Antibody L1Mab-13 Detected Human PD-L1 in Lung Cancers.
Authors: Yamada S, Itai S, Nakamura T, Yanaka M, Chang YW, Suzuki H, Kaneko MK, Kato Y Abstract Programmed cell death ligand-1 (PD-L1) is a type I transmembrane glycoprotein expressed on antigen-presenting cells. It is also expressed in several tumor cells such as melanoma and lung cancer cells. A strong correlation has been reported between human PD-L1 (hPD-L1) expression in tumor cells and negative prognosis in cancer patients. Here, a novel anti-hPD-L1 monoclonal antibody (mAb) L1Mab-13 (IgG1, kappa) was produced using a cell-based immunization and screening (CBIS) method. We investigated hPD-L1 expression in lu...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - April 4, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Soluble Expression in Escherichia coli of a Single-Domain Antibody-Tumor Necrosis Factor α Fusion Protein Specific for Epidermal Growth Factor Receptor.
In this study, we constructed an antibody-cytokine fusion protein (Ia1-TNFα) comprising the single-domain antibody Ia1, which targets epidermal growth factor receptor (EGFR) overexpressed in epithelial tumors and a tumor necrosis factor α (TNFα) domain, which has antitumor activity. Ia1-TNFα was produced in a soluble form by using an Escherichia coli expression system, and after affinity purification of the culture supernatant, an yield of ∼2 mg/L of cell culture was obtained. Gel filtration analysis showed that Ia1-TNFα existed predominantly as a trimer, which is consistent with th...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - February 25, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Detecting Receptor Tyrosine Kinase ROR1 Using a Developed Anti-ROR1 Polyclonal Antibody.
In conclusion, the developed anti-ROR1 PAb can be used as a tool for determining the prognostic value of ROR1 in histopathology of cancer tissues. PMID: 29474159 [PubMed - in process] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - February 25, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generating a Battery of Monoclonal Antibodies Against Firefly Luciferase for Dot Blot Analysis, Western Blot Analysis, and Immunostaining of Cells in Culture and Paraffin Sections.
Authors: Conway TP, Daniels KJ, Park YN, Soll DR Abstract Firefly luciferase (FLuc) is commonly used as a reporter gene PpyLuc1 in bioanalytical assays. We have produced five mouse-derived monoclonal antibodies (mAbs) that recognize FLuc. The mAbs, DSHB-LUC-2, DSHB-LUC-3, DSHB-LUC-9, DSHB-LUC-16, and DSHB-LUC-24, were generated by immunizing mice with purified 6xHIS-tagged FLuc (6xHis-FLuc) in suspension with an adjuvant. All five were validated by dot blots. Four of the mAbs provided strong signals in western blot analysis, and one a weak signal. All five were validated for immunostaining in fixed cell culture. On...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - February 25, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Anti-Semaphorin3Fc Monoclonal Antibody.
Authors: PMID: 29474161 [PubMed - in process] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - February 25, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Synergistic Cytotoxic Effect on Gastric Cancer Cells of an Immunotoxin Cocktail in Which Antibodies Recognize Different Epitopes on CDH17.
Authors: Kusano-Arai O, Iwanari H, Kudo S, Kikuchi C, Yui A, Akiba H, Matsusaka K, Kaneda A, Fukayama M, Tsumoto K, Hamakubo T Abstract Cadherin-17 (CDH17) is highly expressed in gastric cancer and is thus considered to be a good target for antibody therapy. CDH17 is classified as a nonclassical cadherin, in that it is composed of seven extracellular cadherin domains. We generated anti-CDH17 monoclonal antibodies (mAbs) which recognize the extracellular domain of CDH17. Competitive assay using AGS, a gastric cancer cell line, cells revealed that five selected anti-CDH17 mAbs recognize different epitopes on C...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - February 25, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Using a Dihydrofolate Reductase-Based Strategy for Producing the Biosimilar Version of Pertuzumab in CHO-S Cells.
Authors: Ramezani A, Ghaderi A Abstract Targeted therapy using monoclonal antibodies (mAbs) against epidermal growth factor receptor-2 (ErbB2) has been utilized for the treatment of breast cancer, recently. Pertuzumab, one of the anti-ErbB2 mAbs, was approved by FDA in 2012 for the treatment of metastatic breast cancer. The aim of this study was to produce biosimilar version of pertuzumab in Chinese hamster ovary (CHO)-S cell line, and compare its ErbB2-binding and biological activities, with commercial drug, Perjeta. To this end, a dihydrofolate reductase (DHFR)-based strategy was used to produce a CHO-S stable ce...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - February 15, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Preparation, Purification, and Identification of a Monoclonal Antibody Against the C-Terminal Domain of Semaphorin3F.
In this study, our laboratory produced a monoclonal antibody against the C-terminal domain of Sema3F (Sema3Fc mAb) using the hybridoma method, expecting to explore the potential role of the antibody and its application in the detection of Sema3F. The capture enzyme-linked immunosorbent assay (ELISA) method indicated that mAb belonged to the IgM subclass and purified Sema3Fc mAb had a titer of 5.12 × 105 against Sema3Fc by indirect ELISA. In addition, results showed that the Sema3Fc mAb could be applied in such experiments as Western blotting, flow cytometry, immunofluorescence, and immunocytochemical st...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - February 10, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Epitope Mapping of Monoclonal Antibody PMab-52 Against Cat Podoplanin.
In this study, a series of deletion or point mutants of cPDPN were utilized for investigating the binding epitopes of PMab-52 using flow cytometry and Western blotting. The findings of this study revealed that the critical epitopes of platelet aggregation-stimulating domain 4 (PLAG4) of cPDPN are responsible for the binding of PMab-52 to cPDPN. PMID: 29394131 [PubMed - as supplied by publisher] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - February 3, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Immunohistochemical Analysis of Inflammatory Rheumatoid Synovial Tissues Using Anti-Human Podoplanin Monoclonal Antibody Panel.
In this study, we examined PDPN expression in inflammatory synovial tissues using an anti-human PDPN (hPDPN) monoclonal antibody (mAb) panel to select the most useful one for evaluation of synovitis. Synovial tissue samples were obtained from 11 RA patients and 9 osteoarthritis (OA) patients undergoing joint surgery. PDPN-positive cells were immunostained by a panel of PDPN mAbs (NZ-1, LpMab-3, LpMab-7, LpMab-10, LpMab-12, LpMab-13, and LpMab-17), followed by cell grading of inflammation and cell counting of PDPN-positivity by a quantitative analyzer. Immunohistochemistry showed that PDPN was markedly expressed in both mac...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - January 30, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development of an Anti-HER2 Monoclonal Antibody H2Mab-139 Against Colon Cancer.
In this study, we developed a novel anti-HER2 monoclonal antibody (mAb), H2Mab-139 (IgG1, kappa) and investigated it against colon cancers using flow cytometry, western blot, and immunohistochemical analyses. Flow cytometry analysis revealed that H2Mab-139 reacted with colon cancer cell lines, such as Caco-2, HCT-116, HCT-15, HT-29, LS 174T, COLO 201, COLO 205, HCT-8, SW1116, and DLD-1. Although H2Mab-139 strongly reacted with LN229/HER2 cells on the western blot, we did not observe a specific signal for HER2 in colon cancer cell lines. Immunohistochemical analyses revealed sensitive and specific reactions of H2Mab-139 aga...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - January 10, 2018 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Structural Organization of 6B9 Molecule, a Monoclonal Antibody Against Lycopene.
Authors: Petyaev IM, Alekseev KP, Tsibezov VV, Kostina LV, Kozlov AY, Kyle NH, Bashmakov YK Abstract Full cDNA and corresponding amino acid (AA) sequences of 6B9 monoclonal antibody (mAb) against lycopene was obtained using Step-Out RACE technology. Variable (V) and constant (C) regions were identified. The light chain of 6B9 contained 238 AA IgM with the highest level of identity (0.93) to both the anti-VEGF receptor antibody and anti-collagen type II FAb CIIC1. The heavy chain was composed of 634 AA with a high identity (0.9) to the Ig mu chain C region. Potential posttranslational modification regions in both ch...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 22, 2017 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research