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Characterization of mouse‐hamster‐human cross‐reacting antioocyte monoclonal antibodies produced by intrasplenic immunization of mice with 12 zona‐free mouse oocytes
Author(s) -
Jin Meishan,
Johansson Lars,
Sundstrōm Per,
Nilsson B. Ove
Publication year - 1994
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080370411
Subject(s) - zona pellucida , antibody , antigen , biology , hamster , oocyte , immunization , monoclonal antibody , andrology , microbiology and biotechnology , epitope , virology , immunology , embryo , medicine
Several intrasplenic immunizations with batches of ∼15 or ∼30 zona‐free, unfertilized mouse oocytes resulted in 200–300 hybrids, respectively, among which about 20 positive clones were selected from each fusion between splenic plasma cells and SP2/0 myeloma cells. When nonimmunized splenic plasma cells were used, only one antibody, showing weak immunoreaction, was obtained from ∼370 hybrids collected from 2 fusions. From one immunization with a total of 12 zona‐free, unfertilized mouse oocytes, 15 positive clones were selected for further study. Eleven of these 15 antibodies reacted with antigens only in unfertilized oocytes but not in fertilized, pronuclear stage oocytes. Three antibodies, which recognized antigens in paraffin‐embedded oocyte sections, did not label growing ovarian oocytes, indicating that the antibodies were specific to ovulated, unfertilized oocytes. These antibodies did not detect any antigen epitopes in the panel of tissues examined. The molecular weight of one antigen, corresponding to a IgM antibody that is present both in ooplasma and zona pellucida, was ∼116 kDa. Cross‐reactivity to blots of unfertilized zona‐free hamster oocytes was demonstrated by 6 antibodies and to unfertilized human oocytes by 7 antibodies. Three antibodies cross‐reacted with both hamster and human oocytes. The study indicates that the intrasplenic immunization is an appropriate means of raising antibodies against unfertilized, zona‐free mouse oocytes and that the method applied offers an easy way to select antibodies against human oocytes for functional studies. © 1994 Wiley‐Liss, Inc.