A METHOD FOR THE PREPARATION OF A MONOCLONAL ANTIBODY, A MONOCLONAL ANTIBODY, A PHARMACEUTICAL COMPOSITION AND A DIAGNOSTIC REAGENT.

摘要

Method of preparing a monoclonal antibody against the specificity determining part of a cell surface antigen having a constant section and a variable section, where at least a part of said variable section defines a specificity determining part of said surface antigen , consisting of the following steps: 1) injecting a non-human mammal material containing surface antigen, said material being selected from the group consisting of i) whole cells and ii) a fraction of membranes obtained by treating whole cells; 2) isolate a fraction containing B cells from the spleen of said mammal; 3) enriching said cell fraction obtained in step 2 in B cells specific for said cell surface antigen by contacting the cell fraction with support-bound material from cells related to said whole cells, whose related cell material attached to support lacks of said antigen from the cell surface, and separating the B cells bound to the support-bound material from related cells from the enriched cell fraction containing unbound B cells for use in the next step; 4) contacting the cell fraction with material containing antigen from the cell surface bound to support selected from the group consisting of j) whole cells and jj) a fraction of membranes obtained from said whole cells and then separating B cells not bound to said support bound material of B cells bound to said support bound material; 5) subjecting the bound B cells to said support-bound material obtained in the previous step to limiting dilution, followed by clonal expansion; 6) select a B cell clone and immortalize said selected B cell clone using a small scale fusion technique, and 7) select and clone a hybridoma capable of producing antibodies that specifically bind to said cell surface antigen, followed for isolation of a fraction containing monoclonal antibody from the supernatant of said hybridoma.