Studies on monoclonal anti-isotypic and anti-idiotypic antibodies against leukemia and myeloma: VI. Purification and relative affinity of monoclonal antibodies.
In this study, the anti-idiotypic and anti-isotypic antibodies (McAbs) against IgM of the Patient with B chronic lymphocytic leukemia (B-CLL) were purified from hybridoma ascites by n-Octoic acid precipitation method. The purified McAbs have high purify and high antibody activity as evidenced by immunoelectrophoresis, SDS-PAGE and ELISA. Relative affinity of 11 McAbs was measured by using indirect ELISA and double antibody sandwich ELISA method. It was found that relative affinity of various McAbs to the same antigen was different. 11 McAbs could be divided into two groups by analysing their 50% maximum binding. The relative affinity of 4 McAbs in the culture supernatants was consistent with that of McAbs in the purified ascites. Our experimental results provide an important basis for rational application of these McAbs.
Shen
G X
GX
Department of Immunology, Tongji Medical University, Wuhan.
Su
N
N
Wang
X L
XL
Zhu
H F
HF
Zhang
Y
Y
D000888:Antibodies, Anti-Idiotypic / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000915:Antibody Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007075:Immunoglobulin M / Q000276:immunology; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000276:immunology*
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M