High-throughput screening for developability during early-stage antibody discovery using self-interaction nanoparticle spectroscopy.
The discovery of monoclonal antibodies (mAbs) that bind to a particular molecular target is now regarded a routine exercise. However, the successful development of mAbs that (1) express well, (2) elicit a desirable biological effect upon binding, and (3) remain soluble and display low viscosity at high concentrations is often far more challenging. Therefore, high throughput screening assays that assess self-association and aggregation early in the selection process are likely to yield mAbs with superior biophysical properties. Here, we report an improved version of affinity-capture self-interaction nanoparticle spectroscopy (AC-SINS) that is capable of screening large panels of antibodies for their propensity to self-associate. AC-SINS is based on concentrating mAbs from dilute solutions around gold nanoparticles pre-coated with polyclonal capture (e.g., anti-Fc) antibodies. Interactions between immobilized mAbs lead to reduced inter-particle distances and increased plasmon wavelengths (wavelengths of maximum absorbance), which can be readily measured by optical means. This method is attractive because it is compatible with dilute and unpurified mAb solutions that are typical during early antibody discovery. In addition, we have improved multiple aspects of this assay for increased throughput and reproducibility. A data set comprising over 400 mAbs suggests that our modified assay yields self-interaction measurements that are well-correlated with other lower throughput assays such as cross-interaction chromatography. We expect that the simplicity and throughput of our improved AC-SINS method will lead to improved selection of mAbs with excellent biophysical properties during early antibody discovery.
Protein Analytics; Adimab; Lebanon, NH USA.
Yuqi
Y
Liu
Protein Analytics; Adimab; Lebanon, NH USA.
Isabelle
I
Caffry
Center for Biotechnology & Interdisciplinary Studies; Isermann Dept. of Chemical & Biological Engineering; Rensselaer Polytechnic Institute; Troy, NY USA.
Jiemin
J
Wu
Center for Biotechnology & Interdisciplinary Studies; Isermann Dept. of Chemical & Biological Engineering; Rensselaer Polytechnic Institute; Troy, NY USA.
Steven B
SB
Geng
Computational Biology; Adimab; Palo Alto, CA USA.
Tushar
T
Jain
Protein Analytics; Adimab; Lebanon, NH USA.
Tingwan
T
Sun
Protein Analytics; Adimab; Lebanon, NH USA.
Felicia
F
Reid
Protein Analytics; Adimab; Lebanon, NH USA.
Yuan
Y
Cao
Protein Analytics; Adimab; Lebanon, NH USA.
Patricia
P
Estep
Protein Analytics; Adimab; Lebanon, NH USA.
Yao
Y
Yu
Computational Biology; Adimab; Palo Alto, CA USA.
Maximiliano
M
Vásquez
Center for Biotechnology & Interdisciplinary Studies; Isermann Dept. of Chemical & Biological Engineering; Rensselaer Polytechnic Institute; Troy, NY USA.
Peter M
PM
Tessier
Protein Analytics; Adimab; Lebanon, NH USA.
Yingda
Y
Xu
D055910:Antibodies, Immobilized / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000378:metabolism* / Q000627:therapeutic use; D000915:Antibody Affinity; D002478:Cells, Cultured; D055808:Drug Discovery; D006046:Gold / Q000737:chemistry; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D053768:Metal Nanoparticles / Q000737:chemistry / Q000706:statistics & numerical data; D055503:Protein Multimerization; D015203:Reproducibility of Results; D013057:Spectrum Analysis / Q000379:methods*; D020349:Surface Plasmon Resonance
D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D006046:Gold
aggregation; antibody developability; cross-interaction; high-throughput screening; nanoparticle; self-association; self-interaction