Production of nonagglutinating monoclonal antibodies against human red blood cells

Abstract

Nonagglutinating monoclonal antibodies against human red blood cells were produced. A volume of 2 X 107 Ficoll-paque purified red blood cells was used as antigen. Hybrid clones which cause agglutination with red blood cells after adding anti-mouse IgG were selected. Indirect agglutination tests were used for isotype determinations of these antibodies. Eighteen clones producing non-agglutinating antibodies against human red blood cells have been isolated. Among them, sixteen clones produced IgGi(K), while the other two clones produced IgG2b(K)antibodies. Ascites fluid from mouse injected with IgGi(K) type hybridomas were purified, and heavy and light chains of immunoglobulin were identi- fied to have molecular masses of 51 kDa and 25 kDa, respectively.