TY - JOUR

T1 - Effect of experimental conditions on strong biocomplimentary pairing in high-performance monolithic disk affinity chromatography

AU - Ostryanina, Natalia D.

AU - Ilina, Olga V.

AU - Tennikova, Tatiana B.

N1 - Funding Information: We are grateful to Dr M.B. Tennikov for valuable discussions and methodological assistance. The project was partly founded by regional grant of SPb Department of RAS. BIA Separations d.o.o., Ljubljana, Slovenia is greatly acknowledged for providing CIM ® disks.

PY - 2002/4/25

Y1 - 2002/4/25

N2 - The effect of flow-rate on quantitatively determined binding parameters for several biocomplementary pairs in affinity mode high-performance monolithic disk affinity chromatography (HPMDAC) has been investigated using frontal analysis approach. Affinity interactions were evaluated from linearized adsorption isotherms and dynamic dissociation constants of the complexes Kdiss. and the theoretical adsorption capacities Qmax were calculated. HPMDAC isolation of a typical protein trypsin from both buffered solution and artificial mixture as well as biospecific extraction of antibodies against bovine serum albumin and recombinant protein G from such complex mixtures as blood serum and cellular lysate were examined. Immobilized counterparts soybean trypsin inhibitor, bovine serum albumin, and human immunoglobulin G were used in chromatographic experiments. The maximum adsorption capacities obtained at different flow-rates were compared with those determined at static conditions. The dependence of quantitative parameters on the surface density of immobilized ligands has also been explored. Finally, a series of experiments was carried out to evaluate the dependence of dynamic affinity binding on temperature for two complementary pairs.

AB - The effect of flow-rate on quantitatively determined binding parameters for several biocomplementary pairs in affinity mode high-performance monolithic disk affinity chromatography (HPMDAC) has been investigated using frontal analysis approach. Affinity interactions were evaluated from linearized adsorption isotherms and dynamic dissociation constants of the complexes Kdiss. and the theoretical adsorption capacities Qmax were calculated. HPMDAC isolation of a typical protein trypsin from both buffered solution and artificial mixture as well as biospecific extraction of antibodies against bovine serum albumin and recombinant protein G from such complex mixtures as blood serum and cellular lysate were examined. Immobilized counterparts soybean trypsin inhibitor, bovine serum albumin, and human immunoglobulin G were used in chromatographic experiments. The maximum adsorption capacities obtained at different flow-rates were compared with those determined at static conditions. The dependence of quantitative parameters on the surface density of immobilized ligands has also been explored. Finally, a series of experiments was carried out to evaluate the dependence of dynamic affinity binding on temperature for two complementary pairs.

KW - Biocomplenentary pairing

KW - Monolithic disks

UR - http://www.scopus.com/inward/record.url?scp=0037171612&partnerID=8YFLogxK

U2 - 10.1016/S1570-0232(01)00597-9

DO - 10.1016/S1570-0232(01)00597-9

M3 - Article

C2 - 12013242

AN - SCOPUS:0037171612

VL - 770

SP - 35

EP - 43

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SN - 1570-0232

IS - 1-2

ER -