Post-UF/DF Chromatography for Antibody Aggregate Clearance

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Solution Overview

Problem

Existing protein purification methods for biologics and biosimilars from cell culture fail to adequately remove high molecular weight contaminants while maintaining yield, necessitating improved downstream polishing steps to meet stringent purity standards.

Innovation Solution

Incorporating a post-ultrafiltration/diafiltration chromatography step, particularly mixed-mode or cation exchange chromatography, to significantly reduce high molecular weight contaminants in the purification process.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If conventional downstream polishing steps including ultrafiltration/diafiltration are used, then the purification process is efficient and yield is maintained, but high molecular weight contaminants are not adequately removed

Engineering Contradiction:

Improvepurity of target proteinVSAvoidyield of target protein

Core Design Contradiction:

Manufacturing precisionVSLoss of substance

Solution Approach 1:

The purification process is divided into distinct segments: upstream processing, ultrafiltration/diafiltration, and a new post-UF/DF chromatography step. This segmentation allows each step to target specific contaminants at different stages, with the post-UF/DF chromatography specifically addressing high molecular weight contaminants that earlier steps miss, thereby improving purity without compromising overall yield

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention changes the parameter of purification sequence by inserting a chromatography step after ultrafiltration/diafiltration. This parameter change in process sequencing enables the removal of high molecular weight contaminants that would otherwise co-purify with the target protein through conventional steps, achieving both high purity and maintained yield

Inventive Principle:

Principle #35Parameter changes

2Productivity

If high density cell cultures are used to increase yield, then productivity improves, but levels of high molecular weight contaminants increase

Engineering Contradiction:

Improveyield of target proteinVSAvoidhigh molecular weight contaminants

Core Design Contradiction:

ProductivityVSObject-generated harmful factors

Solution Approach 1:

The high molecular weight contaminants generated by high density cell cultures are converted from a harmful byproduct into a targetable substance. The post-UF/DF chromatography step is specifically designed to capture these HMW contaminants, transforming the problem of increased contamination into an opportunity for enhanced purification that allows high density culturing to proceed

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

3Manufacturing precision

If additional chromatography steps are added after ultrafiltration/diafiltration, then high molecular weight contaminants are removed, but process complexity increases

Engineering Contradiction:

Improveclearance of high molecular weight contaminantsVSAvoidnumber of purification steps

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The harmful high molecular weight contaminants are extracted and removed from the purification process at the specific stage after ultrafiltration/diafiltration. This targeted extraction approach removes only the problematic HMW contaminants without requiring complete process redesign, thereby limiting the increase in overall process complexity while achieving the desired purity improvement

Inventive Principle:

Principle #2Taking out (Extraction)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The method achieves a substantial reduction in high molecular weight contaminants by at least 10% while maintaining target protein yield above 58%, conforming to Quality Target Protein Profiles.

Implementation Method 1

contacting the fluid comprising the target protein with a chromatography media

Methodology Applied

Scientific EffectChromatography: Chromatography

Implementation Method 2

at least a mixed-mode chromatography or an ion exchange chromatography (e.g., cation exchange chromatography) step

Methodology Applied

Scientific EffectIon exchange: Ion Exchange

Implementation Method 3

an ultrafiltration step, which may involve diafiltration, a form of ultrafiltration with solvent replenishment

Methodology Applied

Scientific EffectUltrafiltration: Filter (physical)

Data Source

PatentUS20260070941A1Clearance of Aggregates from UF/DF Pools in Downstream Antibody Purification

Publication Date: 2026.03.12 AMGEN INC

AI summary

A method for improving the harvest or purification of a target protein such as a biologic or biosimilar is provided. The method improves conventional harvest/purification methodologies by adding a chromatography step, such as a mixed-mode or ion exchange chromatography step, towards the end of the polishing phase of harvest/purification, after conventional chromatographic polishing steps such as protein A or ion exchange chromatography steps have been completed and the resultant eluate subjected to filtration, such as ultrafiltration/diafiltration. The surprising result of returning to chromatographic polishing after filtration is that all forms of high molecular weight products are reduced, facilitating the purification of target protein sufficient to meet government regulations, such as Quality Target Protein Profiles.