Affinity Chromatography of Human Blood Coagulation Factor VIII on Monoliths with Peptides from a Combinatorial Library

Direct Synthesis of Peptides on Convective Interaction Media Monolithic Columns for Affinity Chromatography

“…35 as achieved by conventional immobilization via epoxy groups. 8 A blank column was prepared by acetylation of the amino groups of a CIM EDA column. The functionality of the supports was tested by injection of a pdFVIII preparation containing the FVIII-vWF complex.…”

“…Previous work had shown that the choice of matrix and spacer is very critical for optimal binding and elution in affinity chromatography. 8,21 The positioning of the peptide and the nature of the support is extremely crucial when peptides are immobilized on a solid phase. Because of steric hindrance and synergism between peptide conformation and the chemistry of the support, the selectivity of a ligand may change with the support.…”

“…2 CIM monolithic supports have been used for the separation of proteins utilizing ion-exchange, reversedphase,hydrophobicinteraction,andaffinitychromatography. 1,[3][4][5][6][7][8] Separation of DNA and oligonucleotides 9,10 and the use of CIM monolithic supports for enzyme biocatalysis 11,12 have also been reported. CIM monolithic supports offer the advantage of high resolution practically unaffected by flow rate, 13,14 low back pressure due to high porosity and short column length, low nonspecific binding of biomolecules, and simple handling.…”

Affinity membrane chromatography for the analysis and purification of proteins

“…Gradient and isocratic elution was investigated and high-performance membrane chromatography experiments were compared with similar ones performed on a conventional packed-bed column. Amw x atschek et al 19 developed an affinity-chromatographic method in which peptides, derived from a combinatorial library, were used as immobilized ligands for the purification of human blood coagulation factor VIII. Trypsin, immobilized on molded macrop-Ž .…”

A Novel Strategy for Proteome-wide Ligand Screening Using Cross-linked Phage Matrices

“…However, the traditional methods to generate affinity ligands such as monoclonal antibodies and synthetic compounds are expensive and time-consuming, which limit the application of affinity technologies (3,4). Fortunately, the advances in phage display technology are providing a highly efficient and inexpensive way to acquire affinity ligands (5)(6)(7). In a phage-display library, ligands are displayed on the virion surface.…”