A novel sensitive assay for functional factor XII based on the generation of kallikrein-C1-inhibitor complexes in factor XII-deficient plasma by glass-bound factor XII.
We developed a very sensitive assay for functional factor XII. This assay is based on the property of glass-bound factor XII to activate prekallikrein (PK) into kallikrein in factor XII-deficient plasma, which is assessed by measuring the formation of kallikrein-C1-inhibitor complexes in this plasma by radioimmunoassay. Incubation of varying amounts of factor XII in glass tubes led to a dose-dependent increase in kallikrein-C1-inhibitor complexes in factor XII-deficient plasma with a lower limit of detection of +/- 20 pg of factor XII. The specificity of the assay for factor XII was demonstrated by experiments with plasmas deficient for factor XII, PK or high molecular weight kininogen (HK) and by incubation of factor XII with Polybrene or with a monoclonal antibody that inhibits the amidolytic and procoagulant activity of factor XII. The high sensitivity of the assay appeared to be due to the inability of C1-inhibitor to inhibit factor XII bound to glass, which resulted in a molar ratio of generated kallikrein to glass-bound factor XII of at least 100:1. This assay for factor XII may be a feasible tool in studies on structural and functional aspects of (recombinant) factor XII species synthesized by cultured cells.
Dors
D M
DM
Central Laboratory, The Netherlands Red Cross Blood Transfusion Service, University of Amsterdam.
Nuijens
J H
JH
Huijbregts
C C
CC
Hack
C E
CE
D003174:Complement C1 Inactivator Proteins / Q000737:chemistry*; D005175:Factor XII Deficiency / Q000097:blood*; D005898:Glass; D006801:Humans; D007610:Kallikreins / Q000737:chemistry*; D011288:Prekallikrein / Q000378:metabolism; D011485:Protein Binding; D012016:Reference Values; D012680:Sensitivity and Specificity
D003174:Complement C1 Inactivator Proteins; D011288:Prekallikrein; D007610:Kallikreins