Affinity-based analytical purification of biotherapeutics for bioprocess monitoring

Patent metadata

Field	Value
Publication number	US-10920182-B2
Application number	US-201715831085-A
Country	US
Kind code	B2
Filing date	Dec 4, 2017
Priority date	Mar 15, 2013
Publication date	Feb 16, 2021
Grant date	Feb 16, 2021

Abstract

The invention as disclosed herein provides a method for purifying a non-antibody protein from solution, comprising a chromatography step wherein the solution is passed over an affinity construct containing an affinity ligand-coupled solid support, wherein the affinity construct is associated with a bioprocess unit operation, and isolating the non-antibody protein from solution.

First claim

The invention claimed is:

1. A method of developing a multi-step bioprocess for producing a non-antibody protein (NAP), the method comprising:
a) processing a NAP comprising an engineered protein or enzyme using a particular bioprocess unit operation within the multi-step bioprocess;
b) purifying the NAP from solution within the multi-step bioprocess by:
i) contacting a heterogeneous solution comprising the NAP with an affinity construct comprising a solid support coupled to an affinity ligand that binds the NAP,
ii) isolating the affinity-purified NAP from the heterogeneous solution, and
iii) determining a critical quality attribute (CQA) of the affinity-purified NAP,
wherein the CQA is a physical, chemical, biological, or microbiological property or characteristic of the NAP selected from the group consisting of product purity, potency, charged isoform profile, post-translational modifications, oxidation, reductions, deamidation, adduct formation, clipped forms, enzymatic cleavage, specific activity, peptide map, dimer content, product aggregation, site specific glycosylation, total glycans, and glycosylation profile;

c) providing a transformed CQA of the affinity-purified NAP by calculating a ratio of the determined CQA of the affinity-purified NAP to a determined CQA of the NAP purified by a multi-step process train purification; and
d) developing the multi-step bioprocess by modifying the particular bioprocess unit operation based on the transformed CQA.

2. The method of claim 1 , wherein the bioprocess unit operation is a bioreactor process, seed train, capture chromatography, intermediate chromatography, filtration, centrifugation, precipitation, flocculation, UV irradiation, or viral inactivation.

3. The method of claim 1 , wherein the transformed CQA of the affinity-purified NAP is equivalent to the NAP produced by a bioprocess unit operation within the multi-step bioprocess.

4. The method of claim 1 , wherein the CQA of the NAP is determined at the following stages within the multi-step bioprocess:
a) immediately upstream of a particular bioprocess unit operation;
b) immediately downstream of the particular bioprocess unit operation;
c) both upstream and downstream of a particular bioprocess unit operation, or
d) within a particular bioprocess unit operation at one or more timepoints.

5. The method of claim 1 , wherein the CQA of the NAP is determined at one or more timepoints within the multi-step bioprocess.

6. The method of claim 1 , wherein the bioprocess is selected from the group consisting of continuous, semi-continuous, and batch.

7. The method of claim 6 , wherein the CQA is measured using a high-throughput or rapid analytical technique.

8. The method of claim 7 , wherein the analytical technique is high-performance liquid chromatography, differential refractometry, fluorescence, ultra-performance liquid chromatography, multi-angle laser light scattering analysis, mass spectroscopy, tandem mass spectroscopy, isoelectric focusing, or differential scanning calorimetry.

9. The method of claim 1 , wherein the NAP is a biotherapeutic drug substance or a commercial biologic.

10. The method of claim 1 , wherein the multi-step bioprocess for the NAP is a commercial or manufacturing process.

11. The method of claim 1 , wherein one or more purification steps are performed by a robot.

12. The method of claim 1 , wherein the ligand of the affinity ligand-coupled based solid support is an antibody.

13. The method of claim 1 , wherein the affinity construct is integrated with the bioprocess in a manner selected from the group consisting of at-line mode, offline mode, and in-line mode.

14. The method of claim 1 , wherein the affinity construct comprises a parameter that is optimized to maximize the quality or purity of the affinity-purified NAP.

Assignees

• Genzyme Corp

Inventors

• Warikoo Veena
• Brower Kevin

Classifications

• C07K1/22 Primary

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• C12Q3/00 Primary

  Condition responsive control processes (apparatus therefor C12M1/36; controlling or regulating in general G05) · CPC title

• C12C3/00 Primary

  Treatment of hops · CPC title

• G01N33/573

  for enzymes or isoenzymes · CPC title