Professor Li Yan's research group has made new progress in the field of rapid quality evaluation of antibody drugs

Recently, Professor Li Yan's research group from the Department of Pharmaceutical Analysis of the School of Pharmacy of Fudan University published a report entitled "Tractable Method for Rapid Quality Assessment of Therapeutic Antibodies in Harvested Cell Culture Fluid based on" in Analytical Chemistry, an authoritative journal in the field of pharmaceutical analysis FcγRIIIa-Immobilized Magnetic Microspheres"
.
This study establishes a new method
for rapid quality evaluation of ADCC-enhanced antibody drugs in samples of engineered cell fermentation broth.

Antibody drugs have become the fastest growing segment of
biotechnology drugs in the world.
Through molecular design and bioengineering, the affinity between antibody molecules and FcγRIIIa receptors on the surface of immune cells can be purposefully improved, and a new generation of antibody drugs
with stronger "antibody-dependent cell-mediated cytotoxicity" (ADCC) effect and better tumor killing effect can be obtained.
However, the traditional quality evaluation scheme for this class of antibody drugs needs to detect multiple key quality attributes including antibody-Fc receptor affinity one by one, which has problems such as
cumbersome operation and long analysis time.

The research team synthesized a FcγRIIIa protein-modified magnetic nanomaterial (_Fe3O4@PDA@PAMAM-FcγRIIIa) with simultaneous coating of polydopamine and dendritic macromolecule PAMAM and applied it as a magnetosolid phase extraction adsorbent for rapid quality evaluation
of engineered cell fermentation broth.

Figure 1.
Workflow of a new method for rapid quality evaluation of antibody drugs

The new Fe3O₄@PDA@PAMAM-Fc γRIIIa material can be directly added to the unpurified engineering cell fermentation broth, selectively binds to ADCC-enhanced antibody components with high affinity, and separates the material from the fermentation broth matrix by applying a magnetic field, which can easily and quickly achieve selective extraction and purification
of high-affinity antibodies 。 The high-affinity antibody eluted from the surface of the material can then be directly analyzed by molecular exclusion chromatography (SEC), and the multi-dimensional key quality attribute information
such as the affinity of the antibody-Fc receptor, the expression of high-affinity antibody and the degree of antibody aggregation in the original fermentation broth sample can be obtained at the same time through the target main peak area and main peak ratio on the chromatogram.
By directly sorting the main peak area, the quality order of antibody drugs in the original fermentation broth sample can be obtained, and then the fermentation broth samples of the antibody drugs with the best quality expression can be quickly screened to guide the subsequent process development
.
This method can complete multi-dimensional quality data detection using only ordinary HPLC, and the operation steps are shortened to less than 20% of the traditional method, and the analysis time is shortened to less than
16% of the traditional method.

Feng Jianan, a 2020 doctoral student at the School of Pharmacy of Fudan University, is the first author of this paper; Professor Li Yan of the School of Pharmacy, Fudan University, is the corresponding author
of this article.
This work was supported
by the National Natural Science Foundation of China.

Original link: https://pubs.
acs.
org/doi/10.
1021/acs.
analchem.
2c01350