التفاصيل البيبلوغرافية
| العنوان: |
High-throughput antibody screening from complex matrices using intact protein electrospray mass spectrometry. |
| المؤلفون: |
Sawyer, William S., Srikumar, Neha, Carver, Joseph, Chu, Phillip Y., Shen, Amy, Ankai Xu, Williams, Ambrose J., Spiess, Christoph, Cong Wu, Yichin Liu, Tran, John C. |
| المصدر: |
Proceedings of the National Academy of Sciences of the United States of America; 5/5/2020, Vol. 117 Issue 18, p1-6, 6p |
| مصطلحات موضوعية: |
COMPLEX matrices, MASS spectrometry, BISPECIFIC antibodies, ANTIBODY formation, IMMUNOGLOBULINS |
| مستخلص: |
Toward the goal of increasing the throughput of high-resolution mass characterization of intact antibodies, we developed a RapidFire-mass spectrometry (MS) assay using electrospray ionization. We achieved unprecedented screening throughput as fast as 15 s/sample, which is an order of magnitude improvement over conventional liquid chromatography (LC)-MS approaches. The screening enabled intact mass determination as accurate as 7 ppm with baseline resolution at the glycoform level for intact antibodies. We utilized this assay to characterize and perform relative quantitation of antibody species from 248 samples of 62 different cell line clones at four time points in 2 h using RapidFire-time-of-flight MS screening. The screening enabled selection of clones with the highest purity of bispecific antibody production and the results significantly correlated with conventional LC-MS results. In addition, analyzing antibodies from a complex plasma sample using affinity-RapidFire-MS was also demonstrated and qualified. In summary, the platform affords high-throughput analyses of antibodies, including bispecific antibodies and potential mispaired side products, in cell culture media, or other complex matrices. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
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