Download PDF
Thromb Haemost 1989; 61(02): 225-229
DOI: 10.1055/s-0038-1646563
Original Article
Schattauer GmbH Stuttgart

Epitope Localization of Anti-Factor VIII Monoclonal Antibodies Determined by Recombinant peptides

Jerry Ware
The Department of Biology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
,
John R Toomey
The Department of Biology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
,
Darrel W Stafford
The Department of Biology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
› Author Affiliations
Further Information

Publication History

Received 01 September 1988

Accepted after revision 06 January 1989

Publication Date:
30 June 2018 (online)

Also available at
 PDF Download  Permissions and Reprints

Summary

In order to define the epitopes recognized by anti-coagulation factor VIII (FVIII) monoclonal antibodies, we have constructed a recombinant DNA epitope library from random fragments of the FVIII cDNA. The characterization of 33 different clones producing recombinant antigens in the expression vector, λgt11, has identified the epitopes for six different anti-FVIII monoclonal antibodies. The antigenic determinant for each antibody was defined by the overlapping or shared DNA sequence of multiple immunoreactive clones. One weak inhibitor of FVIII coagulant activity binds within the gly701 ser750 sequence of the FVIII sequence. An antibody which recognizes the amino-terminus of FVIII heavy-chain (within trp14 - tyr46) does not inhibit FVIII activity. Two non-neutralizing antibodies both map within residues asp807 - ser817 while two other non-neutralizing antibodies bind within lys1673 - pro1688. The general usefulness of this strategy for mapping FVIII antigenic determinants is discussed.

Keywords

Factor VIII - Monoclonal antibodies - Recombinant DNA - Epitope mapping
 

  • References

  • 1 Zimmerman TS, Fulcher CA. Factor VIII procoagulant protein. Clin Haematol 1985; 14 (02) 343-358
    PubMedSearch in Google Scholar
  • 2 Eaton DL, Vehar GA. Factor VIII structure and proteolytic processing. Prog Hemostas Thromb 1986; 8: 47-70
    PubMedSearch in Google Scholar
  • 3 Kane WH, Davie EW. Blood coagulation factor V and VIII: structural and functional similarities and their relationship to hemorrhagic and thrombotic disorders. Blood 1988; 71 (03) 539-555
    PubMedSearch in Google Scholar
  • 4 Gitschier J, Wood WI, Goralka TM, Wion KL, Chen EY, Eaton DH, Vehar GA, Capon DJ, Lawn RM. Characterization of the human factor VIII gene. Nature 1984; 312: 326-330
    CrossrefPubMedSearch in Google Scholar
  • 5 Wood WI, Capon DJ, Simonsen CC, Eaton DL, Gitschier J, Keyt B, Seeburg PH, Smith DH, Hollingshead P, Wion KL, Delwart E, Tuddenham EG D. Expression of active human factor VIII from recombinant DNA clones. Nature 1985; 312: 330-337
    PubMedSearch in Google Scholar
  • 6 Toole JJ, Knopf JL, Wozney JM, Sultzman LA, Bueker JL, Pittman DD, Kaufman RJ, Brown E, Shoemaker C, Orr EC, Amphlett GW, Foster WB. Molecular cloning of a cDNA encoding human antihemophilic factor. Nature 1984; 312: 342-347
    CrossrefPubMedSearch in Google Scholar
  • 7 Truett MA, Blacher R, Burke RL, Caput D, Chu C, Dina D, Hartog K, Kuo CH, Masiarz FR, Merryweather JP, Najarian R, Pachl C, Potter SJ, Puman J, Quiroga M, Rall LB, Randolph A, Urdea MS, Valenzuela P, Dahl HH, Faxalano J, Hansen J, Nordfang O, Ezban M. Characterization of the polypeptide composition of human factor VIII:C and the nucleotide sequence and expression of human kidney cDNA. DNA 1985; 4 (05) 333-349
    CrossrefPubMedSearch in Google Scholar
  • 8 Fulcher CA, Roberts JR, Holland LZ, Zimmerman TS. Human factor VIII procoagulant protein: monoclonal antibodies define precursor-product relationships and functional epitopes. J Clin Invest 1985; 76: 117-124
    CrossrefPubMedSearch in Google Scholar
  • 9 Eaton D, Rodriguez H, Vehar GA. Proteolytic processing of human factor VIII. Correlation of specific cleavages by thrombin, factor Xa, and activated protein C with activation and inactivation of factor VIII coagulant Biochemistry 1986; 25: 505-512
    CrossrefPubMedSearch in Google Scholar
  • 10 Fulcher CA, Roberts JR, Zimmerman TS. Thrombin proteolysis of purified factor VIII procoagulant protein: correlation of activation with generation of a specific polypeptide. Blood 1983; 61 (04) 807-811
    PubMedSearch in Google Scholar
  • 11 Kaufman RJ, Wasley LC, Dorner AJ. Synthesis, processing, and secretion of recombinant human Factor VIII expressed in mammalian cells. J Biol Chem 1988; 263 (13) 6352-62
    PubMedSearch in Google Scholar
  • 12 Pittman DD, Kaufman RJ. Proteolytic requirements for thrombin activation of anti-hemophilic factor (factor VIII). Proc Natl Acad Sci USA 1988; 85 (08) 2429-2433
    CrossrefPubMedSearch in Google Scholar
  • 13 Goodall AH, Meyer D. Registry of monoclonal antibodies to factor VIII and von Willebrand factor. Thromb Haemostas 1985; 54 (04) 878-891
    PubMedSearch in Google Scholar
  • 14 Ware J, Toomey JR, Stafford DW. Localization of a factor VIII- inhibiting antibody epitope to a region between residues 338 and 362 of factor VIII heavy chain. Proc Natl Acad Sci USA 1988; 85 (09) 3165-3169
    CrossrefPubMedSearch in Google Scholar
  • 15 Foster PA, Fulcher CA, Houghten RA, Zimmerman TS. An immunogenic region within amino acid residues val-1670 to glu-1684 of the factor VIII light chain induces antibodies which inhibit binding of factor VIII to von Willebrand factor. J Biol Chem 1988; 263 (11) 5230-5234
    PubMedSearch in Google Scholar
  • 16 Nunberg JH, Rodgers G, Gilbert JH, Snead RM. Method to map antigenic determinants recognized by monoclonal antibodies: localization of a determinant of virus neutralization on the feline leukemia virus envelope protein gp 70. Proc Natl Acad Sci USA 1984; 81: 3675-3679
    CrossrefPubMedSearch in Google Scholar
  • 17 McGraw RA, Frazier D, de Serres M, Reisner H, Stafford DW. Antigenic determinant in human coagulation factor IX: immunological screening and DNA sequence analysis of recombinant phage map a monoclonal to residues 111 through 132 of the zymogen. Blood 1986; 67: 1344-1348
    PubMedSearch in Google Scholar
  • 18 Melgar E, Goldthwait DA. Deoxyribonucleic acid nucleases. J Biol Chem 1968; 243 (17) 4409-4416
    PubMedSearch in Google Scholar
  • 19 Maniatis T, Fritsch EF, Sambrook J. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory; Cold Spring Harbor, NY: 1982. pp 167
    Search in Google Scholar
  • 20 Huynh TV, Young RA, Davies RW. DNA cloning: a practical approach. In: Glover DM. ed IRL Press; Oxford: 1984. pp 49-78
    Search in Google Scholar
  • 21 Benson SA, Taylor RK. A rapid small-scale procedure for isolation of phage lambda DNA. Biotechniques 1984; 2 (03) 126-127
    PubMedSearch in Google Scholar
  • 22 Messing J. New M 13 vectors for cloning. Methods Enzymol 1983; 101: 20-89
    PubMedSearch in Google Scholar
  • 23 Sanger F, Nicklen S, Coulson AR. DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 1977; 74 (12) 5463-5467
    CrossrefPubMedSearch in Google Scholar
  • 24 Biggin MD, Gibson TJ, Hong GF. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 1983; 80: 3963-3965
    CrossrefPubMedSearch in Google Scholar
  • 25 Burke RL, Pachl C, Quiroga M, Rosenberg S, Haigwood N, Nordfang O, Ezaban MJ. The functional domains of coagulation factor VIII: C. J Biol Chem 1986; 261 (27) 12574-12578
    PubMedSearch in Google Scholar
  • 26 Scandella D, Mahoney SD, Mattingly M, Roeder D, Timmons L, Fulcher CA. Epitope mapping of human factor VIII inhibitor antibodies by deletion analysis of factor VIII fragments expressed in Escherichia coli. Proc Natl Acad Sci USA 1988; 85: 6152-6156
    CrossrefPubMedSearch in Google Scholar
  • 27 Lin SW, Dunn JJ, Studier FW, Stafford DW. Expression of human factor IX and its subfragments in Escherichia coli and generation of antibodies to the subfragments. Biochemistry 1987; 25: 505-517
    PubMedSearch in Google Scholar