Extended in vivo half-life of human soluble complement receptor type 1 fused to a serum albumin-binding receptor.
A new approach has been used to extend the T(1/2) of human soluble complement receptor type 1 (sCR1) in rats. The albumin-binding domains B2A3 (BA) and B1A2B2A3 (BABA) from Streptococcal protein G were fused to the carboxyl terminus of sCR1, and the recombinant genes were expressed and amplified in Chinese hamster ovary cells. Western blot analysis and surface plasmon resonance measurements demonstrated the binding of rat serum albumin to both sCR1-BA and sCR1-BABA but not to sCR1. The in vitro complement inhibitory activity of the fusion proteins was shown to be similar to that of sCR1, indicating that neither the albumin-binding domains nor the presence of bovine serum albumin interfere with sCR1 function. Pharmacokinetic analysis showed that the T(1/2) of the distribution phase (T(1/2alpha)) was 3.3, 20.0 and 6.0 min for sCR1, sCR1-BA and sCR1-BABA, respectively. The T(1/2) of the elimination phase (T(1/2beta)) was 103, 297 and 170 min for sCR1, sCR1-BA and sCR1-BABA, respectively. The plasma elimination of sCR1-BA and sCR1-BABA was significantly (P < .05) prolonged as compared to sCR1. The proteins showed similar tissue distribution; at 4-hr postdosing, the highest levels of 125I-radioactivity per gram of tissue were localized in the urine, blood, liver, stomach, and small intestine.
Makrides
S C
SC
Department of Molecular Biology, T Cell Sciences, Inc, Needham, Massachusetts, USA.
Nygren
P A
PA
Andrews
B
B
Ford
P J
PJ
Evans
K S
KS
Hayman
E G
EG
Adari
H
H
Uhlén
M
M
Toth
C A
CA
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D006207:Half-Life; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D017463:Receptors, Complement 3b / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012709:Serum Albumin / Q000378:metabolism*; D012756:Sheep; D014018:Tissue Distribution
D017463:Receptors, Complement 3b; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin