Custom ligand design for biomolecular filtration and purification for bioseperation
OwnerJoseph A. Affholterinventor
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Abstract
The instant invention relates to a method of isolating a bio-active molecule of interest from a biological system using affinity separation. The affinity separation ligand for the bio-active molecule of interest is selected by directed molecular evolution to customize the properties of the interaction between the bio-active molecule of interest and the affinity separation ligand thereby facilitating isolation of the bio-active molecule of interest. The invention also relates to compositions comprising an affinity separation ligand bound to a solid support wherein the separation ligand is selected by the method of directed molecular evolution.
Claims20
A method of isolating an affinity ligand for affinity separation of a molecule of interest comprising the following steps:
- a) selecting a starting ligand;
- b) altering the starting ligand using a process of reiterative diversification;
- c) isolating the altered ligand of b);
- d) screening the isolated ligand of c) for the presence of a desired phenotype;
- e) optionally repeating any of steps b), c) and d);
- f) chemically linking the ligand of d) or e) to a solid support.
A composition comprising an affinity ligand chemically linked to a solid support wherein said affinity ligand is obtained by a process of reiterative diversification.
A method of isolating a molecule of interest using a custom designed affinity ligand comprising the following steps:
- a) obtaining a genetic library which encodes a candidate affinity ligand for the molecule of interest;
- b) altering the genetic library using a method of reiterative diversification such that the affinity ligand expresses a desired phenotype;
- c) expressing the altered genetic library of b);
- d) screening the altered library for an affinity ligand for the molecule of interest wherein said altered affinity ligand expresses the desired phenotype;
- e) optionally repeating any of b), c) and/or d);
- f) isolating the affinity ligand after any of steps c), d), or e);
- g) chemically linking the isolated affinity ligand of f) to a solid support;
- h) contacting the affinity ligand linked to the solid support with the molecule of interest under a first set of conditions such that the molecule of interest binds to the affinity ligand of g);
- i) altering the first set of conditions of h) such that the molecule of interest separates from the affinity ligand linked to the solid support.
Dependencies are inferred from the claim text.