![Fig. 4. Epitope mapping of LE2E9. / (A) Structure of circulating fVIII. Each domain of fVIII is identified by a letter in upper case followed by a numeral. The small acidic regions are indicated by a1, a2, and a3, respectively. The copper ion involved in the non-covalent association of the A1 and A3 domains is indicated. (B) Recombinant fVIII fragments labeled with [35S]methionine and expressed in reticulocyte lysates were incubated for 2 hours at 4°C with LE2E9 bound to Protein A Sepharose. After washing, bound material was eluted and analyzed by SDS-PAGE, followed by autoradiography. Recombinant fVIII fragments are indicated by open bars. The amino- and carboxy-terminal extremities of fragments are as indicated. Control experiments were carried out with a human monoclonal antibody, BO2C11, directed toward the fVIII C2 domain19 and a rabbit polyclonal IgG antibody, aA3, directed toward amino acid residues 1797-1815 of the fVIII A3 domain The experiments were repeated at least 3 times with comparable results.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/95/1/10.1182_blood.v95.1.156/5/bloo00150004x.jpeg?Expires=1736047354&Signature=OTeUCM2M-zacKeoA4gnLnlZ8YqKn-35RiJNmdU-YI36h5L~GaIXAsw8EwuglclZN5UjxfkXx1IsW5MvNGDoAEyJXZnQ0XqmmGk8UOzGmvo~bKBpKTqLQ6nZZ5lyosWafw~pU0OiiZlkak-vg33tq7o-z3kU4EnjQ0KHEPdjBUMhwl7DnMP-Jdt7CQ0AtpuHzECwTBABf2wn96Lns5abYdf3VEGghUCrFg0DBA8UfY7ntfM0yrvFXUp2Y7YcAov1t2Wb7sqDzYrqUir-wXU6Xls6zbZB~-B8c733vwOPq4qe6fv8fY2c3AXsth8LC~SCLVxBEq4-zzI1XEsGmZCCQuw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Epitope mapping of LE2E9.
(A) Structure of circulating fVIII. Each domain of fVIII is identified by a letter in upper case followed by a numeral. The small acidic regions are indicated by a1, a2, and a3, respectively. The copper ion involved in the non-covalent association of the A1 and A3 domains is indicated. (B) Recombinant fVIII fragments labeled with [35S]methionine and expressed in reticulocyte lysates were incubated for 2 hours at 4°C with LE2E9 bound to Protein A Sepharose. After washing, bound material was eluted and analyzed by SDS-PAGE, followed by autoradiography. Recombinant fVIII fragments are indicated by open bars. The amino- and carboxy-terminal extremities of fragments are as indicated. Control experiments were carried out with a human monoclonal antibody, BO2C11, directed toward the fVIII C2 domain19 and a rabbit polyclonal IgG antibody, aA3, directed toward amino acid residues 1797-1815 of the fVIII A3 domain The experiments were repeated at least 3 times with comparable results.
