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Development of a droplet digital PCR for pertussis toxin locus copy number determination in a genetically-modified Bordetella pertussis strain
Biologicals. 2023 May 5;82:101683. doi: 10.1016/j.biologicals.2023.101683. Online ahead of print.ABSTRACTTo improve pertussis toxin (PT) yield in B. pertussis strains for vaccine production a genetically-engineered strain (gdPT 191-134 strain) with a second copy of the genetically detoxified PT (gdPT) locus was developed. The consistency of the production and genetic stability of the strain when used for vaccine production must be established. We developed two simplex ddPCR assays with PCR systems for ptxA, the target gene present in two copies, and pgm, the reference gene present as a single copy. The ddPCR assay had suff...
Source: Biologicals : Journal of the International Association of Biological Standardization - May 7, 2023 Category: Biotechnology Authors: Rapha ël Esson St éphanie Falque Eric Abachin Steve George Nolwenn Nougarede Source Type: research

Development and validation of a spectrophotometric method for quantification of residual cyclodextrin (DIMEB; Heptakis) in pertussis antigens
We report here development and validation of a spectrophotometric method for estimation of DIMEB in different product matrices of pertussis vaccine antigens i.e. Filamentous haemagglutinin (FHA), Pertactin (PRN) and Pertussis toxin (PT). The detection is based on characteristic reaction of hydrolyzed sugars derivatives from DIMEB i.e., furfural derivatives with anthrone reagent to form colored complexes which could be quantified at 625 nm. Method showed excellent linearity with correlation coefficient (R2) > 0.995 over the concentration of 5.0-80.0 μg. LOD and LOQ of 1.47 μg and 4.46 μg respectively was reported. The...
Source: Biologicals : Journal of the International Association of Biological Standardization - February 15, 2023 Category: Biotechnology Authors: Bharat Shinde Dadasaheb Patil Vinod Nandre Manish Gautam Pooja Doshi Sunil Gairola Source Type: research

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