title: Standardization and validation of Vero cell assay for potency estimation of diphtheria antitoxin serum.
abstract: Diphtheria toxin has the capacity to block protein synthesis in cultured mammalian cells, and thus causing cell death. This capacity of diphtheria toxin was utilized for in-vitro neutralization test to determine antibody titer, using Vero cells, which have been found to be susceptible to diphtheria toxin. In the present study, a Vero cell assay was standardized and validated for potency estimation of diphtheria antitoxin serum (DATS). The results obtained by Vero cell assay were compared with in-vivo biological assay. High degree of correlation (+0.98) was found between in-vivo biological assay and in-vitro Vero cell assay. The assay has also been found to be effective in determining the rising antibody titer in the equines inducted in DATS production. The present study indicated that although biological assays hold the key for final potency estimations till date but in the future scenario in-vitro Vero cell assay may be a good alternative to in-vivo biological assay.
authors.affiliation: Central Research Institute, Kasauli, Himachal Pradesh 173204, India. sunilk63@rediffmail.com
authors.forename: Sunil
authors.initials: S
authors.lastname: Kumar
authors.forename: Sarika
authors.initials: S
authors.lastname: Kanwar
authors.forename: Vivek
authors.initials: V
authors.lastname: Bansal
authors.forename: Rakesh
authors.initials: R
authors.lastname: Sehgal
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D002522:Chlorocebus aethiops; D004166:Diphtheria Antitoxin / Q000032:analysis* / Q000276:immunology / Q000627:therapeutic use; D005260:Female; D006168:Guinea Pigs; D006736:Horses; D007106:Immune Sera / Q000032:analysis / Q000276:immunology; D008297:Male; D008954:Models, Biological; D009500:Neutralization Tests / Q000379:methods* / Q000592:standards*; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors; D014709:Vero Cells
chemical_list: D004166:Diphtheria Antitoxin; D007106:Immune Sera