Derivation and Application of Monoclonal Antibodies Recognizing Several Epitopes on Bovine Serum Albumin.

The establishment of a highly sensitive ELISA for detecting bovine serum albumin (BSA) based on a specific pair of monoclonal antibodies (mAb) and its application in vaccine quality control

“…The detectable concentration of existing mAb sandwich ELISA was only 10 µg/L. 15 In our assay, the limit of detection is 0.38 µg/L which is 10-fold more sensitive than existing mAb sandwich ELISA, and which is comparable to the currently used commercial pAb ELISA (0.25 µg/L). The cutoff value of BSA content in commercial pAb ELISA is 32 µg/L, which in our assay is 40 µg/L.…”

“…15 In our assay, we performed epitope-analysis studies with SPR, by which antibodies were first separated into different groups. From 11 hybridomas, we selected 4 for the subsequent pairwise interaction analysis, and finally the best pairwise antibodies were chosen (the data obtained for other interfering antibodies is not shown).…”

“…However, in some subsequent studies employing serum-free Hymedium 606, some hybridomas were also difficult to culture. 15 Later, it was reported that goat serum could be used as an alternative to fetal bovine serum in biomedical research. 17 Thus, by modifying the hybridoma technique, we tested both horse and goat serum in RPMI 1640 medium as the complete medium to In the present study, the BSA sandwich ELISA assay was designed to provide a simple, precise, and highly sensitive method to detect BSA contamination.…”

“…However, the cutoff value in existing mAb sandwich ELISA is 1,500 µg/L. 15 According to WHO requirement, the upper limit of BSA content in vaccines is 50 µg/L. It is very difficult to use the existing mAb sandwich ELISA according to WHO guidelines.…”

“…15 Here we report a highly sensitive and stable mAb sandwich ELISA, based on the development of a panel of mAbs against BSA that possess high affinity and that recognize different epitopes on the BSA molecule.…”