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Abstract

A method of producing a purified lipid-binding peptide which can bind to phospholipids at one or more amphipatic alpha-helical peptide regions. The method includes providing a gene coding for the peptide, and introducing the gene in expressible, heterologous form in a suitable expression system capable of synthesizing a mixture of peptides which includes the lipid-binding peptide. Addition of either endogenous or exogenous lipids to the peptide mixture forms a low-density lipopeptide complex composed of lipid and the lipid-binding peptide, and this complex can be separated easily from nonlipid-binding peptides in the peptide on the basis of its size and/or density. The method is intended particularly for scaled-up production of purified human apolipoproteins and their alpha-helical lipid-binding regions. Also disclosed are related methods for producing recombinant apolipoproteins, therapeutic lipopeptide compositions, and a stabilized lipid emulsion for nutritional therapy. Further disclosed are methods for expressing apolipoproteins or lipid-binding segments thereof in bacterial, yeast and mammalian cell expression systems, and methods for purifying lipid binding proteins, including fused recombinant proteins.

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First Claim