Non-viral adeno-associated virus-based platform for stable expression of antibody combination therapeutics.
Antibody combination therapeutics (ACTs) are polyvalent biopharmaceuticals that are uniquely suited for the control of complex diseases, including antibiotic resistant infectious diseases, autoimmune disorders and cancers. However, ACTs also represent a distinct manufacturing challenge because the independent manufacture and subsequent mixing of monoclonal antibodies quickly becomes cost prohibitive as more complex mixtures are envisioned. We have developed a virus-free recombinant protein expression platform based on adeno-associated viral (AAV) elements that is capable of rapid and consistent production of complex antibody mixtures in a single batch format. Using both multiplexed immunoassays and cation exchange (CIEX) chromatography, cell culture supernatants generated using our system were assessed for stability of expression and ratios of the component antibodies over time. Cultures expressing combinations of three to ten antibodies maintained consistent expression levels and stable ratios of component antibodies for at least 60 days. Cultures showed remarkable reproducibility following cell banking, and AAV-based cultures showed higher stability and productivity than non-AAV based cultures. Therefore, this non-viral AAV-based expression platform represents a predictable, reproducible, quick and cost effective method to manufacture or quickly produce for preclinical testing recombinant antibody combination therapies and other recombinant protein mixtures.
1Excelimmune, Inc.; Lexington, MA USA.
Gwendolyn M
GM
Wilmes
1Excelimmune, Inc.; Lexington, MA USA.
Kimberly L
KL
Carey
1Excelimmune, Inc.; Lexington, MA USA.
Stuart W
SW
Hicks
1Excelimmune, Inc.; Lexington, MA USA.
Hugh H
HH
Russell
1Excelimmune, Inc.; Lexington, MA USA.
Jesse A
JA
Stevenson
1Excelimmune, Inc.; Lexington, MA USA.
Paulina
P
Kocjan
1Excelimmune, Inc.; Lexington, MA USA.
Stephen R
SR
Lutz
1Excelimmune, Inc.; Lexington, MA USA.
Rachel S
RS
Quesenberry
1Excelimmune, Inc.; Lexington, MA USA.
Sergey V
SV
Shulga-Morskoy
1Excelimmune, Inc.; Lexington, MA USA.
Megan E
ME
Lewis
1Excelimmune, Inc.; Lexington, MA USA.
Ethan
E
Clark
1Excelimmune, Inc.; Lexington, MA USA.
Violetta
V
Medik
1Excelimmune, Inc.; Lexington, MA USA.
Anthony B
AB
Cooper
1Excelimmune, Inc.; Lexington, MA USA.
Elizabeth E
EE
Reczek
D002460:Cell Line; D002852:Chromatography, Ion Exchange / Q000379:methods; D018920:Coculture Techniques; D000229:Dependovirus; D004359:Drug Therapy, Combination; D015870:Gene Expression; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D011994:Recombinant Proteins; D057127:Single-Chain Antibodies
adeno-associated virus; antibody cocktails; antibody combination therapeutics; antibody mixtures; biologics expression; monoclonal antibodies