Development of a peptidomimetic ligand for efficient isolation and purification of factor VIII via affinity chromatography.
Hemophilia A, one of the most severe bleeding disorders, results from an inherited deficiency of factor VIII (FVIII) function. Treatment by injection of FVIII has been a common procedure for decades. Nevertheless, the production and purification of FVIII remains a challenging task. Current procedures using immunoaffinity chromatography are expensive and suffer from the instability of the applied antibody ligands, which elute along with the product and contaminate it. Recently, FVIII was purified by use of octapeptide ligands, but their low protease-resistance limits their application. We here report the systematic rational and combinatorial optimization procedure that allowed us to transfer the octapeptide ligands into a small peptidomimetic. This compound is the smallest ligand known for separation of such a large protein (330 kDa). It not only binds and purifies FVIII with high efficiency but also is stable, protease-resistant, and cheap to produce in preparative scale. Hence it offers a valuable alternative to antibody-based purification procedures.
Knör
Sebastian
S
Center of Integrated Protein Science Munich, Department Chemie, Lehrstuhl II für Organische Chemie, Technische Universität München, Lichtenbergstrasse 4, Garching D-85747, Germany.
Khrenov
Alexey V
AV
Laufer
Burkhardt
B
Saenko
Evgueni L
EL
Hauser
Charlotte A E
CA
Kessler
Horst
H
D019943:Amino Acid Substitution; D002846:Chromatography, Affinity / Q000379:methods; D004355:Drug Stability; D005169:Factor VIII / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D007210:Indoleacetic Acids / Q000138:chemical synthesis* / Q000737:chemistry; D008024:Ligands; D018716:Molecular Mimicry; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry; D011108:Polymers; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D044967:Serum; D013237:Stereoisomerism
C523904:5-(1-(1-carboxy-2-mercaptoethylcarbamoyl)-2-(4-hydroxyphenyl)ethylamino)-4-(2-1H-indol-3-ylacetylamino)pentanoic acid; D007210:Indoleacetic Acids; D008024:Ligands; D009842:Oligopeptides; D011108:Polymers; D011994:Recombinant Proteins; D005169:Factor VIII; D010447:Peptide Hydrolases