High-throughput screening for developability during early-stage antibody discovery using self-interaction nanoparticle spectroscopy
| Autor: | Tingwan Sun, Jiemin Wu, Steven B. Geng, Felicia Reid, Yuan Cao, Peter M. Tessier, Yingda Xu, Maximiliano Vásquez, Tushar Jain, Isabelle Caffry, Yuqi Liu, Patricia Estep, Yao Yu |
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| Rok vydání: | 2014 |
| Předmět: |
medicine.drug_class
High-throughput screening Immunology Antibody Affinity Nanoparticle Metal Nanoparticles Monoclonal antibody Report High-Throughput Screening Assays Drug Discovery medicine Immunology and Allergy Humans Throughput (business) Cells Cultured biology Chemistry Spectrum Analysis Antibodies Monoclonal Reproducibility of Results Surface Plasmon Resonance Combinatorial chemistry Biochemistry Polyclonal antibodies Colloidal gold biology.protein Gold Immunotherapy Antibody Protein Multimerization Antibodies Immobilized |
| Zdroj: | mAbs. 6(2) |
| ISSN: | 1942-0870 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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