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S J Tarnowski

Publications and source records attributed to S J Tarnowski.

12 recordsLinked to original sources
PubMed1987

Rapid dual-column chromatographic assay for recombinant leukocyte interferon alpha-2.

A rapid dual-column chromatographic assay for determining recombinant leukocyte interferon alpha-2 in complex mixtures is described. The assay relies on the use of a high-performance monoclonal antibody affinity column connected in tandem with a reversed-phase column. The high specificity and selectivity of these columns permits the quantitation of subcomponent species, such as interferon oligomers that may be present in assay samples. The assay has a limit of sensitivity equal to 1 microgram/g over a range of 1-20 micrograms/g. The precision of the assay was estimated to be about 5%.

Chromatography, Affinity↗
PubMed1985

Analysis of different forms of recombinant human leukocyte interferons and synthetic fragments by high-performance liquid chromatography.

Analytical high-performance liquid chromatography (HPLC) conditions are reported for the evaluation of recombinant human interferon monomers: recombinant human leukocyte interferon A (rIFN-alpha A), rIFN-alpha D, and the hybrid rIFN-alpha A1-62/D64-166. The two monomeric forms of rIFN-alpha A (slow-migrating monomer and fast-migrating monomer) were also resolved by HPLC. Conditions are reported for the HPLC separation of oligomers (dimers and trimers) of rIFN-alpha A. The synthesis and analytical HPLC of the carboxy-terminus fragment, corresponding to IFN-alpha D (140-166), and a series of analogues comprising the IFN-alpha A (105-125) region is reported. The syntheses were accomplished by the solid-phase peptide synthesis procedure and the products were purified by preparative HPLC.

Amino Acid Sequence↗
PubMed1980

Coenzyme A-synthesizing protein complex of Saccharomyces cerevisiae.

The coenzyme A-synthesizing protein complex (CoA-SPC) is a multienzyme complex of Saccharomyces cerevisiae (Bakers' yeast), which has a molecular weight in excess of 200,000 as determined by Sephadex G-200 column chromatography. This multienzyme complex, which is insoluble in the crude yeast cell lysate, has been purified 229-fold. A cellular component of the yeast cell lysate, referred to as t-Factor, with a molecular weight of 400-1000 and chloride ion are involved in the solubilization of CoA-SPC. The CoA-SPC requires L-cysteine, D-pantothenic acid and ATP as substrates. The terminal CoA-SPC-bound intermediate is dephospho-CoA, which is subsequently phosphorylated and released from the complex as CoA. The sequence of reactions for the synthesis of CoA by the CoA-SPC differs significantly from those previously proposed for other systems. It could be that the reaction sequence is unique for the yeast cell.

Adenosine Triphosphate↗
PubMed1980

Alternate procedure for the preparation of the coenzyme A-synthesizing protein complex of Bakers' yeast.

The coenzymes A-synthesizing protein complex (CoA-SPC) of Bakers' yeast synthesizes coenzyme A in an in vitro system from the precursors ATP, D-pantothenic acid and L-cysteine. CoA-SPC has been produced on a small scale by freezing Bakers' yeast cells in a mixture of diethyl ether and solid CO2, followed by a thawing period, and subsequent removal of the diethyl ether by vacuum. The resulting yeast lysate was then stirred for 18 h in the presence of t-Factor to solubilize CoA-SPC. When a greater quantity of CoA-SPC was needed, the danger associated with the use of a large volume of diethyl ether was apparent. Therefore, the freezing step involving diethyl ether and solid CO2 has been replaced by a process of slowly drying fresh Bakers' yeast until approximately 34% of the initial weight of the yeast remained as dry solids. The yeast solids were ground to further disrupt the cell wall and membrane structure. The grinding step was followed by rehydration of the dry yeast solids with deionized H2O and stirring the rehydrated yeast for 18 h to solubilize CoA-SPC.

Coenzyme A↗