Large-scale purification of active platelet integrin glycoprotein IIb-IIIa.
Glycoprotein IIb-IIIa is an abundant platelet receptor of the integrin family that plays a primary role in platelet aggregation. It exists on the platelet surface predominantly in a resting or inactive conformation that is converted to an active binding competent conformation upon platelet activation. There is much interest in studying the difference between active and inactive GP IIb-IIIa, developing therapeutic agents targeted towards GP IIb-IIIa and developing diagnostic assays for antibodies that recognize epitopes on GP IIb-IIIa. We present here the development of a large-scale process for purifying active GP IIb-IIIa from human platelets. The procedure results in 25mg batch sizes of high purity and activity. Additionally, the effects of detergent concentration and impurities such as IgG on ELISA assays are examined.
Department of Biotechnology, Experimental Station, Bristol-Myers Squibb Pharma Company, Wilmington, DE 19880, USA.
Milton C
MC
Hillman
Nina I
NI
Zolotarjova
Denis R
DR
Patrick
Denise D
DD
McCabe
Keqiang
K
Shen
Jeanne I
JI
Corman
Jeffrey T
JT
Billheimer
Dietmar A
DA
Seiffert
Gregory F
GF
Hollis
Richard
R
Wynn
D001792:Blood Platelets / Q000737:chemistry*; D015153:Blotting, Western; D002457:Cell Extracts; D002846:Chromatography, Affinity; D003208:Concanavalin A / Q000378:metabolism; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D009842:Oligopeptides / Q000378:metabolism; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000302:isolation & purification* / Q000378:metabolism; D013997:Time Factors
D002457:Cell Extracts; D003902:Detergents; D007074:Immunoglobulin G; D009842:Oligopeptides; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D003208:Concanavalin A; C046617:arginyl-glycyl-aspartyl-serine