Cassette baculovirus vectors for the production of chimeric, humanized, or human antibodies in insect cells.
Plasmid cassette-transfer vectors pBHuC chi and pBHuC gamma l have been designed which enable the construction of recombinant baculoviruses directing the co-expression of complete immunoglobulin in insect cells. We describe the application of these vectors for the expression of a human/mouse chimeric monoclonal antibody of potential immunosuppressive clinical value derived from a mouse anti-human CD29 monoclonal antibody (Mu-K20). The chimeric K20 light and heavy chains produced in sf9 insect cells were correctly processed and assembled into a normal immunoglobulin which is secreted into the culture medium of infected cells. The chimeric mAb Ch-K20-sf9 reproduces in vitro the functional properties of the parental mouse K20, including affinity and inhibition of lymphocyte proliferation. These results demonstrate that the baculovirus/insect cell expression system is suitable for the expression of fully active monoclonal antibodies of therapeutic value. Our generic cassette approach makes this system a very flexible and convenient one for the rapid production of either chimeric, humanized or human mAb with heavy and light chains of any isotype.
Poul
M A
MA
Laboratoire d'ImmunoGénétique Moléculaire, CNRS, Universités Montpellier I et II, France.
Cerutti
M
M
Chaabihi
H
H
Ticchioni
M
M
Deramoudt
F X
FX
Bernard
A
A
Devauchelle
G
G
Kaczorek
M
M
Lefranc
M P
MP
D000818:Animals; D000906:Antibodies / Q000235:genetics*; D000918:Antibody Specificity; D015703:Antigens, CD / Q000378:metabolism; D001681:Biological Assay; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D019012:Integrin beta1; D016023:Integrins / Q000378:metabolism; D051379:Mice; D016716:PC12 Cells; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D018411:Spodoptera
D000906:Antibodies; D015703:Antigens, CD; D019012:Integrin beta1; D016023:Integrins; D011993:Recombinant Fusion Proteins