Figure 3
Figure 3. VWF multimers and degradation products postinfusion of rVWF-rFVIII. Dotted horizontal lines indicate the zone where bands of ULMW multimers appear. Due to limitations in space, it was not possible to run all samples taken from a single patient during the PK study on 1 gel. Therefore, to show the distribution of multimers and fragments over time, it was necessary to combine different gels into 1 picture. Vertical lines have been inserted to indicate repositioned gel lanes. (A) Typical VWF multimer pattern (upper gels; low-resolution agarose [1% Seakem]; samples adjusted to VWF:Ag content) and fragments cleaved by ADAMTS13 (lower gels; SDS-PAGE followed by immunoblot with polyclonal anti-VWF antibody; samples were applied undiluted) postinfusion of rVWF-rFVIII. (B) Typical VWF multimer pattern (upper gels; low-resolution agarose [1% Seakem]; samples adjusted to VWF:Ag content) and fragments cleaved by ADAMTS13 (lower gels; patient plasma diluted to 0.05 VWF:Ag U/mL with VWD plasma; 0.025 mU VWF:Ag per lane applied) postinfusion of pdVWF-pdFVIII. (C) Subject #21, with specific pretreatment nonneutralizing anti-VWF–binding antibodies (upper gels: low-resolution agarose [1% Seakem]; samples adjusted to VWF:Ag content; lower gels: SDS-PAGE followed by immunoblot with polyclonal anti-VWF antibody; samples were applied undiluted). (D) Subject #23, with specific pretreatment nonneutralizing anti-VWF–binding antibodies.

VWF multimers and degradation products postinfusion of rVWF-rFVIII. Dotted horizontal lines indicate the zone where bands of ULMW multimers appear. Due to limitations in space, it was not possible to run all samples taken from a single patient during the PK study on 1 gel. Therefore, to show the distribution of multimers and fragments over time, it was necessary to combine different gels into 1 picture. Vertical lines have been inserted to indicate repositioned gel lanes. (A) Typical VWF multimer pattern (upper gels; low-resolution agarose [1% Seakem]; samples adjusted to VWF:Ag content) and fragments cleaved by ADAMTS13 (lower gels; SDS-PAGE followed by immunoblot with polyclonal anti-VWF antibody; samples were applied undiluted) postinfusion of rVWF-rFVIII. (B) Typical VWF multimer pattern (upper gels; low-resolution agarose [1% Seakem]; samples adjusted to VWF:Ag content) and fragments cleaved by ADAMTS13 (lower gels; patient plasma diluted to 0.05 VWF:Ag U/mL with VWD plasma; 0.025 mU VWF:Ag per lane applied) postinfusion of pdVWF-pdFVIII. (C) Subject #21, with specific pretreatment nonneutralizing anti-VWF–binding antibodies (upper gels: low-resolution agarose [1% Seakem]; samples adjusted to VWF:Ag content; lower gels: SDS-PAGE followed by immunoblot with polyclonal anti-VWF antibody; samples were applied undiluted). (D) Subject #23, with specific pretreatment nonneutralizing anti-VWF–binding antibodies.

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