Abstract
Monoclonal antibodies derived from ten hybrid cell clones, generated against porcine zona pellucida gave strong immunofluorescence with zona but the pattern varied from patchy, thin rim to heavy precipitation type of rim. Five of the 6 monoclonals studied prevented the binding of the porcine epididymal sperm to homologous oocytesin vitro, whereas the sixth one was partially effective. All of the 6 monoclonale of this batch inhibited the lysis of zonae by proteolytic enzymes even at dilutions up to 1 × 10−3. Three of the four monoclonals prepared in a subsequent batch gave strong immunofluorescent reactions and had high titres as determined by enzyme immunoassay. These monoclonals did not, however, protect the zonae against lysis by proteolytic enzymes. These properties are suggestive of the heterogeneity of the antigenic determinants in zona and emphasize the employment of appropriate bioassays for screening and selection of bioeffective antibodies.
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Abbreviations
- EIA:
-
Enzyme immunoassay
- ZP:
-
zona pellucida
- MoAb:
-
monoclonal antibodies
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- BSA:
-
bovine serum albumin
- PBS:
-
0.01 M phosphate buffered-saline pH 7.2
- PA-HRP:
-
protein A-horse radish peroxidase
- FITC:
-
fluorescein isothiocyanate
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Bamezai, A.K., Bhattacharya, A., Band, A.H. et al. Diversity of antigenic determinants of porcine zona pellucida revealed by monoclonal antibodies. J Biosci 7, 105–113 (1985). https://doi.org/10.1007/BF02703579
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DOI: https://doi.org/10.1007/BF02703579