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Semin Thromb Hemost 2002; 28(3): 297-308
DOI: 10.1055/s-2002-32666
Copyright © 2002 by Thieme Medical Publishers, Inc., 333 Seventh Avenue, New York, NY 10001, USA. Tel.: +1(212) 584-4662

Analysis of Samples from Patients Treated with ReFacto® for the Presence of Anti-SQ-Peptide Specific Antibodies

Margareta Wikén, Katarina Sjöberg
  • Department of Biopharmaceutical Development, Biovitrum AB, Stockholm, Sweden
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Publication History

Publication Date:
04 July 2002 (online)

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ABSTRACT

ReFacto® lacks the major part of the B-domain, except for 14 amino acid residues from the N-terminal and C-terminal ends, which are linked into a 14 amino acid SQ-peptide tail in the C-terminal end of the 90 kD heavy chain, thereby introducing a new peptide sequence in the factor VIII (FVIII) molecule. Therefore, samples from some of the patients enrolled in the ReFacto clinical trials were analyzed for the emergence of anti-SQ-peptide antibodies in addition to determinations for inhibitors by Bethesda assay and for anti-FVIII antibodies by enzyme-linked immunosorbent assay (ELISA). An anti-peptide ELISA was developed using six partially overlapping 15-16 mer peptides covering the SQ-linker region. Results from ReFacto-treated anti-FVIII antibody-positive or -negative patients were compared with those for normal healthy adults and patients treated with other FVIII products. The pattern was compared with that for SQ-peptide specific monoclonal antibodies, each recognizing three or four overlapping SQ-peptides. Antibodies recognizing mainly one of the SQ-peptides were found in samples in some individuals from all investigated groups. This means that the anti-SQ reactivity found in patients treated with ReFacto is not induced by the SQ-sequence of the molecule but rather by some other agent, giving rise to antibodies cross-reactive with SQ-peptides.

KEYWORDS

ReFactor® - SQ-peptide - continuous epitope - immunogenicity - human antibodies

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