Human antibodies for immunotherapy development generated via a human B cell hybridoma technology.
Current strategies for the production of therapeutic mAbs include the use of mammalian cell systems to recombinantly produce Abs derived from mice bearing human Ig transgenes, humanization of rodent Abs, or phage libraries. Generation of hybridomas secreting human mAbs has been previously reported; however, this approach has not been fully exploited for immunotherapy development. We previously reported the use of transient regulation of cellular DNA mismatch repair processes to enhance traits (e.g., affinity and titers) of mAb-producing cell lines, including hybridomas. We reasoned that this process, named morphogenics, could be used to improve suboptimal hybridoma cells generated by means of ex vivo immunization and immortalization of antigen-specific human B cells for therapeutic Ab development. Here we present a platform process that combines hybridoma and morphogenics technologies for the generation of fully human mAbs specific for disease-associated human antigens. We were able to generate hybridoma lines secreting mAbs with high binding specificity and biological activity. One mAb with strong neutralizing activity against human granulocyte-macrophage colony-stimulating factor was identified that is now considered for preclinical development for autoimmune disease indications. Moreover, these hybridoma cells have proven suitable for genetic optimization using the morphogenics process and have shown potential for large-scale manufacturing.
Li
Jian
J
Morphotek, Inc., 210 Welsh Pool Road, Exton, PA 19341, USA.
Sai
Tao
T
Berger
Marc
M
Chao
Qimin
Q
Davidson
Diane
D
Deshmukh
Gaurav
G
Drozdowski
Brian
B
Ebel
Wolfgang
W
Harley
Stephen
S
Henry
Marianne
M
Jacob
Sara
S
Kline
Brad
B
Lazo
Ella
E
Rotella
Frank
F
Routhier
Eric
E
Rudolph
Kathryn
K
Sage
Jeaneen
J
Simon
Paul
P
Yao
Jun
J
Zhou
Yuhong
Y
Kavuru
Mani
M
Bonfield
Tracey
T
Thomassen
Mary Jane
MJ
Sass
Philip M
PM
Nicolaides
Nicholas C
NC
Grasso
Luigi
L
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000627:therapeutic use*; D000941:Antigens / Q000235:genetics; D001327:Autoimmune Diseases / Q000276:immunology / Q000628:therapy; D001402:B-Lymphocytes / Q000276:immunology*; D020137:Base Pair Mismatch; D002478:Cells, Cultured; D004260:DNA Repair; D000939:Epitopes / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D017578:Immunoglobulin Class Switching; D007167:Immunotherapy / Q000379:methods*; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests
D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D016178:Granulocyte-Macrophage Colony-Stimulating Factor