| Abstract: |
IgM molecules were purified by the use of anti-IgM antibody-coupled Sepharose from the culture supernatant of an Epstein-Barr-virus-transformed lymphoblastoid cell line, MP1, that secretes alloantibodies possessing HLA-DQw2 specificity as defined by the cytotoxicity assay. The obtained IgM preparation was labeled with radioactive iodine-125I and fractionated by gel filtration. It contained pentameric IgM and smaller oligomeric IgMs. When tested by the direct cellular binding assay against a panel of HLA-typed cell lines, they all showed the DR3 and DR7 association pattern characteristic of DQw2. A weak but significant binding was detected for DR1, DR6, and DR9. On isoelectrofocusing, MP1 pentameric IgM gave a restricted banding pattern comparable to monoclonal IgM obtained from a patient with Waldenström's syndrome. Moreover, the pattern was identical to that of IgM purified from the culture supernatant of a defined hybrid clone, 162, that was generated by fusing MP1 cells with heteromyeloma D33 cells. The target class II molecules showed the dimeric structure that conforms to DQw2 molecules. © 1989. |
| Keywords: |
human cell; cells, cultured; monoclonal antibody; antibodies, monoclonal; cell culture; protein purification; cell transformation; radioisotope; epstein barr virus; antibody formation; cell transformation, viral; herpesvirus 4, human; electrophoresis, polyacrylamide gel; immunoglobulin m; cytotoxicity tests, immunologic; alloantibody; hla antibody; lymphoblast; human; priority journal; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; hla-dq antigens; hla d antigen
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