Fundamental and practical studies on high-performance liquid affinity chromatography of biopolymers with novel stationary phases

Thesis/Dissertation · Wed Jan 01 04:00:00 EST 1992

OSTI ID:6974723

Bacolod, M D

Rigid microparticulate stationary phases having surface-bound metal chelating functions were developed and evaluated in high performance metal chelate affinity chromatography of proteins. Silica- and polystyrene-divinylbenzene-based metal chelate sorbents were produced in wide pore and in non-porous type of column packings. A major effort has been placed on development of non-porous highly crosslinked polystyrene-divinylbenzene (PSDVB). These PSDVB microparticles were produced by a two-step swelling polymerization, and exhibited excellent mechanical strength over a wide range of flow-rates and composition used in high performance liquid chromatography (HPLC). Simple and reproducible hydrophilic coatings were developed for the surface modification of hydrophobic PSDVB supports. A tetradentate metal chelating ligand, ethylenediamine-N, N[prime]-diacetic acid (EDDA), was covalently bound to the surface of the various supports. Sorbents having iminodiacetic acid (IDA) metal chelating functions were also evaluated. The hydrophilic character and surface coverage of various stationary phases were assessed chromatographically. Studies concerning the effects of eluent pH as well as the nature and concentration of salts on retention and selectivity with different metal chelate stationary phases having various immobilized metal ions were carried out. Elution schemes were developed for rapid separation of proteins in metal chelate affinity chromatography. EDDA stationary phases in metal forms can be viewed as complementary to IDA stationary phases since they afforded different selectivity and retentivity toward proteins. Hydrophilic PSDVB could be functionalized with IDA or EDDA metal chelating ligands or lectins. The non-porous metal chelate stationary phases afforded rapid separation of proteins by the development of multiple gradient systems, which permitted higher column peak capacity, enabling the separation of a greater number of proteins in a single chromatographic run.

• Research Organization: Oklahoma State Univ., Stillwater, OK (United States)
• OSTI ID: 6974723
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
400105* -- Separation Procedures
400201 -- Chemical & Physicochemical Properties
CHELATES
CHEMICAL PROPERTIES
CHROMATOGRAPHY
COMPLEXES
ION EXCHANGE CHROMATOGRAPHY
ION EXCHANGE MATERIALS
LIQUID COLUMN CHROMATOGRAPHY
MATERIALS
ORGANIC COMPOUNDS
ORGANIC ION EXCHANGERS
ORGANIC POLYMERS
POLYMERS
POLYOLEFINS
POLYSTYRENE-DVB
PROTEINS
SEPARATION PROCESSES