Purification and Immunochemical Characterization of Human Plasma Factor XIII

Human plasma Factor XIII (F. XIII) complex is composed of two types of noncovalently linked polypeptide chains termed a and b; only the a chain possesses catalytic potential. Platelet Factor XIII is comprised solely of a chains which are identical to those found in plasma. In this study platelets were utilized as a source of unbound a chains to characterize F. XIII (a chain)-binding activity in plasma and its subfractions. Upon exclusion chromatography of unheated plasma or of an unheated ammonium sulfate (20% sat.) subfraction, F. XIII activity emerged in a peak corresponding to a mol. wt. of < 500,000 (region 1). If these samples had first been heated at 60° to precipitate fibrinogen, F. XIII was eluted in a peak corresponding to a mol wt. of about 300,000 (region 2). Chromatography of the platelet zymogen alone resulted in a F. XIII peak corresponding in position to that of monomeric a chain (mol. wt. 80,000, region 3).Exclusion chromatography of the unheated ammonium sulfate fraction yielded, in addition to the F. XIII peak in region 1, a protein peak (peak II) in region 2 which contained no F. XIII. When peak II was mixed with platelet F. XIII, and again subjected to exclusion chromatography, the platelet F. XIII peak shifted from its expected position in region 3 and emerged instead in region 2 ; this behavior demonstrated F. XIII (a chain)-binding activity within peak II. The same chromatographic shift was observed in mixtures of platelet F. XIII and normal plasma or that from a patient with congenital F. XIII (a chain) deficiency. Immunochemical analyses of chromatographic fractions indicated that a chain-binding was due to complexing of a chains with freely circulating b chains. Since a chains and b chains have different biosynthetic sites we conclude that b chains serve as an extracellular F. XIII (a chain)-binding protein.Supported by NHLI grant HL-11409.

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Composition and structure of plasma lipoproteins. Isolation, composition, and immunochemical characterization of low density lipoprotein subfractions of human plasma

Biochemistry ◽

10.1021/bi00813a004 ◽

1970 ◽

Vol 9 (11) ◽

pp. 2244-2252 ◽

Cited By ~ 135

Author(s):

Diana M. Lee ◽

Petar Alaupovic

Keyword(s):

Human Plasma ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Plasma Lipoproteins ◽

Low Density ◽

Lipoprotein Subfractions ◽

Immunochemical Characterization ◽

Composition And Structure

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Characterization of the kinetic pathway for fibrin promotion of .alpha.-thrombin-catalyzed activation of plasma factor XIII

Biochemistry ◽

10.1021/bi00218a008 ◽

1991 ◽

Vol 30 (4) ◽

pp. 934-941 ◽

Cited By ~ 55

Author(s):

Michael C. Naski ◽

Laszlo Lorand ◽

Jules A. Shafer

Keyword(s):

Factor Xiii ◽

Plasma Factor

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Characterization of a Monoclonal Antibody Directed Against the Carboxyl-Terminus of Human Factor XIII

Thrombosis and Haemostasis ◽

10.1055/s-0038-1642385 ◽

1994 ◽

Vol 71 (01) ◽

pp. 062-067 ◽

Cited By ~ 2

Author(s):

Yeqing Song ◽

Stephen M Taubenfeld ◽

DeQuan Sheng ◽

Gary R Matsueda

Keyword(s):

Monoclonal Antibody ◽

Factor Xiii ◽

Human Factor ◽

Carboxyl Terminus ◽

Heat Denaturation ◽

Structural Element ◽

Assay Format ◽

Plasma Factor ◽

Carboxyl Terminal

SummaryBy deriving an anti-peptide monoclonal antibody, mAb 7A4, we characterized the relatively unstudied carboxyl-terminal end of the α-chain of human factor XIII, the plasma transglutaminase. MAb 7A4 was directed against the last eight amino acids (Gln-Ile-Gln-Arg-Arg-Pro-Ser-Met) and bound with a dissociation constant of 3.4 × 10−8 M. In a solid assay format, mAb 7A4 bound equally well to factor XIII obtained from human plasma, platelets or placenta. However, in a solution-phase assay format, the epitope was largely unavailable but could be readily exposed by heat denaturation. Tmmunoblotting showed that this epitope is conserved among all species of plasma factor XIII tested except rabbit suggesting that the carboxyl-terminus might be an important structural element. Other competitive binding experiments with synthetic peptides as inhibitors pointed toward the final carboxyl-terminal amino acid, Met-731, as an immunochemically important determinant. This was used advantageously to confirm the finding that the caiboxyl-leiminal Mel-731 is largely absent from placental factor XIII (1) as compared to platelet or plasma factor XIII.

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The Activation of Plasma Factor XIII with the Snake Venom Enzymes Ancrod and Batroxobin Marajoensis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1651479 ◽

1977 ◽

Vol 38 (02) ◽

pp. 0438-0446 ◽

Cited By ~ 9

Author(s):

M Walter ◽

D Nyman ◽

V Krajnc ◽

F Duckert

Keyword(s):

Human Plasma ◽

Snake Venom ◽

Factor Xiii ◽

Test System ◽

Plasma Factor

SummaryThe snake venom enzymes Ancrod and Batroxobin marajoensis are able to activate human plasma factor XIII as shown by the formation of the γ-dimers. The concentration of γ-dimers increases with the concentration of the activating enzymes. Factor XIII activated by Ancrod or Batroxobin marajoensis is, however, unable to catalyse the incorporation of the amine dansylcadaverine into casein. The partially activated factor XIII is therefore not demonstrable by means of the artificial test system. This factor XIII loses little activity and remains activable by thrombin.

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A Pasteurised Concentrate of Human Plasma Factor XIII for Therapeutic Use

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661573 ◽

1986 ◽

Vol 55 (03) ◽

pp. 402-405 ◽

Cited By ~ 5

Author(s):

L Winkelman ◽

G E Sims ◽

M E Haddon ◽

D R Evans ◽

J K Smith

Keyword(s):

Human Plasma ◽

Intravenous Injection ◽

Plasma Proteins ◽

Factor Xiii ◽

Sodium Citrate ◽

Therapeutic Use ◽

Freeze Dried ◽

Plasma Factor ◽

Ethanol Fraction

SummaryA therapeutic concentrate of factor XIII containing both A and B sub-units has been prepared from 300 kg pools of human plasma. The process starts from a cold-ethanol fraction from cryoprecipitate supernatant and therefore does not interfere with the recovery of other clinically valuable plasma proteins. Factor XIII is purified approximately 600-fold from plasma by precipitation with sodium citrate and by the removal of fibrinogen by brief heating. The product has been pasteurised in sorbitol solution to inactivate blood-borne viruses, ultrafiltered to remove sorbitol, adsorbed with bentonite and freeze-dried in a formulation meeting requirements for intravenous injection.

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