Clinical Significance of Antibodies to Bovine and Human Thrombin and Factor V After Surgical Use of Bovine Thrombin

“…Factor V inhibitors are relatively rare and have been most frequently reported in the postsurgical setting and/or in association with aminoglycoside antibiotics (3). Several recent reports, however, have described patients developing Factor V inhibitors after exposure to topical bovine thrombin preparations during cardiovascular or neurosurgical procedures (4,5). In one report, the topical thrombin preparation was shown to contain bovine Factor V, which may have served as the immunogen in that patient (4).…”

Section: Introductionmentioning

confidence: 99%

“…Factor Va procoagulant activity was measured using a chromogenic assay, as previously described ( 16). Briefly, the Factor V sample was first activated with RVV-V (3.75 Ag/ml) and added to 50 mM Tris HCl, pH 7.9, 175 mM NaCl, 5 Aliquots ofthe reaction mixture were transferred to microtiter plate wells, and the amount of thrombin generated during the reaction was determined by the addition of0.2 mM S2238 (D-Phe-L-pipecolyl-Arg-p-nitroanilide) and measuring the change in absorbance at 405 nm using a V.,, microtiter plate reader (Molecular Devices, Inc., Menlo Park, CA). Standard curves were prepared using purified thrombin.…”

Section: Introductionmentioning

confidence: 99%

Characterization of an acquired inhibitor to coagulation factor V. Antibody binding to the second C-type domain of factor V inhibits the binding of factor V to phosphatidylserine and neutralizes procoagulant activity.

Ortel

Quinn-Allen²,

Charles³

et al. 1992

J. Clin. Invest.

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Coagulation Factor V is an essential component of the prothrombinase complex, which activates the zymogen prothrombin to thrombin. A patient was described who developed a Factor V inhibitor that neutralized the procoagulant activity of Factor V and resulted in a fatal hemorrhagic diathesis (Coots, M. C., A. F. Muhleman, and H. I. Glueck. 1978. Am. J. Hematol. 4:193-206). This inhibitor was shown to be an IgG antibody that bound to the light chain of Factor V. Using a series of light chain deletion mutants, we have found that this antibody binds to the second C-type domain of the light chain. Both inhibitor IgG and Fab fragments rapidly neutralized the procoagulant activity ofFactor Va, implying that the neutralization resulted from specific binding to the C2 domain. We have previously demonstrated that deletion of the C2 domain results in loss of procoagulant activity, as well as loss of phosphatidylserine-specific binding. Confirming these results, both inhibitor IgG and Fab fragments interfered with phosphatidylserinespecific binding of Factor V. Conversely, preincubation of Factor Va with procoagulant phospholipids protected the cofactor from inactivation by the inhibitor. Our results suggest that this inhibitor neutralizes the procoagulant activity of Factor Va by interfering with the C2-mediated interaction with phospholipid surfaces, thereby disrupting formation of the prothrombinase complex. (J. Clin. Invest. 1992. 90:2340-2347

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“…[9][10][11][12] It is clear from these results that despite the presence of detectable amounts of bovine factor Va antigens in crude thrombin, this contaminant in crude thrombin failed to elicit the generation of antibodies against factor Va light chain in rabbits. This may be due to a difference in the immune responses among species (ie, unlike the immune response to bovine thrombin, rabbit and human may have a different immune response to the bovine factor V/ Va antigens).…”

Section: Discussionmentioning

confidence: 99%

Reduced Immunogenic Potential of Membrane Filtered Bovine Thrombin Preparations for Hemostatic Application

Zhu

Hoppensteadt

Wahi

et al. 2007

Clin Appl Thromb Hemost

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on the reduction of trace contaminants such as the bovine factor Va/Va light chain fragments to an undetectable level in thrombin 4B. In order to compare the relative immunogenic potential, bovine crude thrombin and its purified versions, 4A and 4B preparations, were administered intravenously to individual groups of rabbits on day 0 and day 21 using standard immunologic protocols. Antiserum was drawn from each rabbit on day 30 and the pooled antisera were purified to obtain the IgGs using protein G affinity columns. The antibody profile for each of these IgGs was determined by using specific human and bovine coagulation factors such as prothrombin, thrombin, factor Xa, factor VIIa, and factor Va fragment. In addition, bovine crude thrombin as well as 4A and 4B preparations were also profiled in cross immunoblotting studies. Western blotting results suggest thrombin 4B IgG has the most selective binding with thrombin antigen, and thrombin 4B has the least immunogenic potential among the 3 thrombin preparations tested. Furthermore, neither cross-reactivity A lthough topical bovine thrombin preparations have been widely used for hemostatic purposes in clinical practice, there is concern that exposure to bovine thrombin can result in the development of antibodies, usually against factor V/Va, which can lead to hemostatic abnormalities. It is thought that coagulopathy following exposure to topical thrombin may be attributed to the impurities in bovine thrombin preparations and that purer preparations of bovine thrombin might be less immunogenic. Utilizing newer methods including a membrane filtration step, bovine crude thrombin is further purified into thrombin 4A and 4B preparations which exhibit a higher specific activity and are devoid of some of the protein contaminants. A previous study has reported There is concern that exposure to bovine thrombin can result in the development of antibodies, usually against factor V/Va, which can lead to hemostatic abnormalities. It is thought that purer preparations of bovine thrombin might be less immunogenic. Utilizing newer methods including a membrane filtration step, bovine crude thrombin is further purified into thrombin 4A and 4B preparations which exhibit a higher specific activity and are devoid of some of the protein contaminants. Bovine crude thrombin and its purified versions were administered intravenously to individual groups of rabbits using standard immunologic protocols.Antiserum was drawn from each rabbit and the pooled antisera were purified to obtain the IgGs using protein G affinity columns. The results suggest that the reported purification process, including filtration, resulted in the removal of most of the antigens found in crude thrombin, and that none of these preparations generated any detectable antibodies against bovine factor V related antigens.

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Clinical Significance of Antibodies to Bovine and Human Thrombin and Factor V After Surgical Use of Bovine Thrombin

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Tissue adhesives in ocular surgery

Tissue adhesives in ocular surgery

Characterization of an acquired inhibitor to coagulation factor V. Antibody binding to the second C-type domain of factor V inhibits the binding of factor V to phosphatidylserine and neutralizes procoagulant activity.

Reduced Immunogenic Potential of Membrane Filtered Bovine Thrombin Preparations for Hemostatic Application

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