Chromatography ligand comprising domain C from
View Patent ↗The present invention relates to a chromatography ligand, which comprises Domain C from Staphylococcus protein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIP) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates to a process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.
1. A process of isolating one or more target compounds, which process comprises contacting a liquid comprising said compound(s) with a chromatography matrix; allowing said compound(s) to adsorb to ligands present on the matrix, wherein said ligands comprise at least one of a Staphylococcus protein A (SpA) Domain C sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NO 1 or SEQ ID NO: 2, and, optionally, eluting said compound(s) by the passing across said matrix of a liquid that releases them from ligands, wherein said ligands are coupled to a matrix/carrier and wherein said ligands further comprise a terminal coupling group, said group preferably comprising at least one nitrogen and/or sulphur atom(s), wherein said process further includes an alkaline regeneration step and is carried out at least 80 times.
2. A process of isolating one or more target compounds, which process comprises contacting a liquid comprising said compound(s) with a chromatography matrix; allowing said compound(s) to adsorb to ligands present on the matrix, wherein said ligands are multimers comprising at least two Staphylococcus protein A (SpA) Domain C units; and, optionally, eluting said compound(s) by the passing across said matrix of a liquid that releases them from ligands, wherein the Domain C sequence comprises a sequence of amino acids selected from the group consisting of SEQ ID NO 1 or SEQ ID NO: 2, wherein said process further includes an alkaline regeneration step and is carried out at least 80 times.
3. The process of claim 1 , wherein the target compound is a protein.
4. The process of claim 1 , wherein the alkaline regeneration step comprises incubation in a NaOH solution.
5. The process of claim 4 , wherein the NaOH solution has a concentration of at least 0.5 M.
6. The process of claim 1 , wherein the process is carried out at least 200 times.
7. The process of claim 1 , wherein the target compound is a Fab fragment.
8. The process of claim 2 , wherein the alkaline regeneration step comprises incubation in a NaOH solution.
9. The process of claim 8 , wherein the NaOH solution has a concentration of at least 0.5 M.
10. The process of claim 9 , wherein the process is carried out at least 200 times.
11. The process of claim 2 , wherein the target compound is a Fab fragment.
12. The process of claim 1 , wherein the target compound is an antibody.