PRODUCTION OF MONOCLONAL RECOMBINANT ANTIBODIES WITHOUT THE USE OF HYBRIDOMAS BY IIN VITRO SPLEEN FRAGMENT CULTURE COMBINED WITH ISOTHERMAL SELF-SUSTAINED SEQUENCE REPLICATION OF RNA
摘要
2138009 9411507 PCTABS00032 The invention is directed to a method for producing a recombinant protein having a preselected binding affinity and specificity for a given antigen. Generally, the method combines spleen fragment culture for the in vitro somatic mutation of B cells in response to antigen, followed by preferential amplification of RNA encoding heavy and light chains of desired antibodies produced by those B cells. The amplified RNA is then converted to DNA and incorporated into expression vectors; transfected host cells then express the antibody chains or a single chain antibody having preselected characteristics. Also disclosed are generic degenerate primer pools for the efficient amplification of RNA encoding mouse IgG.
申请公布号
CA2138009(A1)
申请公布日期
1994.05.26
申请号
CA19932138009
申请日期
1993.11.19
申请人
GINGERAS, THOMAS R.;KLINMAN, NORMAN R.;STILLMAN, CATHY A.;LINTON, PHYLLIS-JEAN;DECKER, DEBRA J.;BIERY, MATTHEW C.
发明人
GINGERAS, THOMAS R.;KLINMAN, NORMAN R.;STILLMAN, CATHY A.;LINTON, PHYLLIS-JEAN;DECKER, DEBRA J.;BIERY, MATTHEW C.