Prometic Biosciences Inc.

American National Red Cross

Baldev Singh Baines

Jason Richard Betley

Steven James Burton

James Christopher Pearson

David John Hammond

John Curling

Christopher Bryant

Dwun-Hou Chen

Timothy Keith Hayes

Claudia Hildegard Kuhn

The invention discloses methods for sequential protein isolation and purification from a biological sample by affinity chromatography. Affinity chromatography is conducted using ligands or ligand support complexes that selectively and specifically bind to proteins in the biological sample. The ligands or ligand support complexes were contacted sequentially in a predetermined order with the biological sample to allow each ligand or ligand-support complex to sequentially bind a protein from the biological sample.

Sequential protein isolation and purification schemes by affinity chromatography

For the separation, removal, isolation, purification, characterization, identification or quantification of plasminogen or a protein that is a plasminogen analogue, an affinity adsorbent is used that is a compound of formula (II) wherein one X is N and the other is N, C&#x2014;Cl or C&#x2014;CN; A is a support matrix, optionally linked to the triazine ring by a spacer; Z is O, S or N&#x2014;R and R is H, Calkyl, Chydroxyalkyl, benzyl or &bgr;-phenylethyl; B is an optionally substituted hydrocarbon linkage containing from 1 to 10 carbon atoms; D is H, OH or a primary amino, secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidino or amidino group; or B-D is &#x2014;CHCOOH&#x2014;(CH)&#x2014;NH; and q is 2 to 6.

Affinity adsorbents for plasminogen

For the separation, removal, isolation, purification, characterization, identification or quantification of Factor VIII, von Willebrand's Factor or a protein that is a analogue of either, an affinity adsorbent is used that is a compound of formula (II) wherein one X is N and the other is N, C&#x2014;Cl or C&#x2014;CN; A is a support matrix, optionally linked to the triazine ring by a spacer; Y is O, S or NR; Z is O, S or N&#x2014;R; Rand Rare each H, Calkyl, Chydroxyalkyl, benzyl or &bgr;-phenylethyl; B and W are each an optionally substituted hydrocarbon linkage containing from 1 to 10 carbon atoms; D is H, OH or a primary amino, secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidino or amidino group; or B-D is &#x2014;CHCOOH&#x2014;(CH)&#x2014;NH; and Ris a group bearing a positive charge at neutral pH.

Affinity adsorbents for Factor VIII and von Willebrand's Factor

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Cambridge, United Kingdom

Baldev Singh Baines