Development of sandwich enzyme-linked immunosorbent assay for determination of tetanus toxoid concentration.
According to the recommendation of the World Health Organization (WHO), the use of an in vivo test for measuring of the potency of tetanus toxoid vaccine (TTdV) is still unavoidable, but the establishment of a convenient in vitro test would significantly improve the work in this field. A sandwich enzyme-linked immunosorbent assay (sELISA) was developed for a rapid and sensitive quantification of tetanus toxoid (TTd). We produced four monoclonal antibodies (MAbs) designated 41, 51, 62, and 71 that reacted with TTd and recognized different antigenic determinants on TTd. We also used two of these antibodies for developing a sELISA, with MoAb 71 as an immobilized and MoAb 51 as a capture antibody. The measurement range of this assay was from 31-1000 ng/mL and the minimum detection limit for TTd was 31 ng/mL. This high sensitivity of this sELISA and its good reproducibility suggest that the developed method could be reliably used to estimate the concentration of TTd, which could be easily extrapolated to the estimation of vaccine potency.
Seatović
Sasa
S
Institute of Immunology and Virology-Torlak, Belgrade, Yugoslavia.
Inić-Kanada
Aleksandra
A
Stojanović
Marijana
M
Zivković
Irena
I
Jankov
Ratko M
RM
Dimitrijević
Ljiljana
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013745:Tetanus Toxoid / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D013745:Tetanus Toxoid