A molecular approach for isolating high‐affinity Fab fragments that are useful in blood group serology

Czerwiński, Marcin; Krop‐Watorek, Anna; Siegel, Don L.; Spitalnik, Steven L.

doi:10.1046/j.1537-2995.1999.39499235667.x

Cited by 19 publications

(28 citation statements)

References 42 publications

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“…To evaluate their affinity and specificity in more detail, soluble F(ab) were produced 14 . In addition, we used light‐chain shuffling to produce novel high‐affinity F(ab) that agglutinated RBCs using an antiglobulin method 15 …”

Section: Gpa Polymorphisms In Antigens M and Nmentioning

confidence: 99%

Construction of dimeric F(ab) useful in blood group serology

et al. 2002

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Section: Gpa Polymorphisms In Antigens M and Nmentioning

confidence: 99%

Construction of dimeric F(ab) useful in blood group serology

et al. 2002

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“…Although the phage display technique can be used in multiple areas of basic research and in numerous clinical applications, it may prove particularly useful in transfusion medicine 7,11,12 . A critical step toward improving the practice of transfusion medicine would be a better understanding of the immune response after blood transfusion or during pregnancy.…”

Section: Applications Of Phage Display In Transfusion Medicinementioning

confidence: 99%

The phage display technique and transfusion medicine

Jarolím¹

2001

Transfusion

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“…Wild-type mice can be actively immunized with hGPA 53 and many mouse mAbs are available. [12][13][14][15]18,19,53,[60][61][62][63] This type of blood group polymorphism (ie, the presence or absence of an entire protein) also occurs in humans: En(a Ϫ ) RBCs lack hGPA, S Ϫ s Ϫ U Ϫ RBCs lack human glycophorin B, and "Rh-negative" RBCs lack the Rh(D) protein. …”

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confidence: 99%

Mouse models of IgG- and IgM-mediated hemolysis

Schirmer

Song

Baliff

et al. 2006

Blood

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Well-characterized mouse models of alloimmune antibody-mediated hemolysis would provide a valuable approach for gaining greater insight into the pathophysiology of hemolytic transfusion reactions. To this end, mouse red blood cells (mRBCs) from human glycophorin A transgenic (hGPA-Tg) donor mice were transfused into non-Tg recipients that had been passively immunized with IgG or IgM hGPA-specific monoclonal antibodies (mAbs). In this novel murine "blood group system," mRBCs from hGPA-Tg IntroductionHemolytic transfusion reactions (HTRs) are dangerous complications of blood transfusions. IgM-mediated HTRs (IgM-HTRs) occur acutely, are usually due to ABO incompatibility, typically cause intravascular hemolysis, and can lead to shock, renal failure, coagulopathy, and death. 1 Although infrequent, IgM-HTRs have a high mortality rate. IgG-mediated HTRs (IgG-HTRs) are more common, but usually less severe, than IgM-HTRs; they can occur acutely or be delayed, typically cause extravascular hemolysis, and occasionally result in renal failure, coagulopathy, and death. 2 Symptomatic immune-mediated red blood cell (RBC) destruction also occurs in autoimmune hemolytic anemia (AIHA), 3 blood group incompatible transplantation, 4 and immune thrombocytopenic purpura (ITP) treated with Rh-immune globulin. 5 Although various different modalities are used to treat HTRs, there is no definitive evidence regarding efficacy. Given the sporadic nature of HTRs, designing human trials to evaluate treatment options is difficult. Therefore, to study the mechanisms underlying HTRs, to evaluate the efficacy of existing treatments, and to develop new treatments, a relevant, inexpensive, and tractable animal model is required. 6 A mouse model would be ideal for this purpose, 7 given the abundance of reagents and relevant knockout (KO) and transgenic (Tg) animals. Although mouse blood group polymorphisms exist, 8 and transfused incompatible mouse RBC (mRBCs) are rapidly cleared, 9,10 no one has exploited these findings to develop a model of HTRs. The availability of a Tg mouse expressing human glycophorin A (hGPA; gene symbol designation, GYPA) on mRBCs 11 offers an approach using a well-described glycoprotein target and well-characterized reagents.We describe initial studies validating mouse models of IgG-and IgM-mediated alloimmune hemolysis. Thus, mRBCs from hGPA-Tg donors were transfused into non-Tg recipients that were passively immunized with IgG or IgM hGPA-specific monoclonal antibodies (mAbs). The clearance of the transfused mRBCs was quantified and the roles of complement and Fc␥ receptors were evaluated. Materials and methods AntibodiesPurified mAbs NaM10-2H12 (IgG3), NaM26-3F4 (IgG1), and NaM10-6G4 (IgG2a), which recognize peptide epitopes on the M and N forms of hGPA, 12 were purchased from EFS (Nantes, France). Hybridomas producing mAb 6A7, an IgG1 specific for a sialic acid-dependent epitope on M-type hGPA, 13-15 10F7, an IgG1 recognizing a nonpolymorphic epitope on hGPA, 13,14 and J11d, a rat IgM recognizing CD24 on mRBCs, 16,17...

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A molecular approach for isolating high‐affinity Fab fragments that are useful in blood group serology

Cited by 19 publications

References 42 publications

Construction of dimeric F(ab) useful in blood group serology

Construction of dimeric F(ab) useful in blood group serology

The phage display technique and transfusion medicine

Mouse models of IgG- and IgM-mediated hemolysis

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