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Immunology and Infection

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Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries

18.5K Views

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12:55 min

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January 17th, 2015

DOI :

10.3791/51492-v

January 17th, 2015

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Shane Miersch¹^,², Zhijian Li¹^,², Rachel Hanna¹^,², Megan E. McLaughlin¹^,², Michael Hornsby¹^,³, Tet Matsuguchi¹^,³, Marcin Paduch¹^,⁴, Annika Sääf¹^,⁴, Jim Wells¹^,³, Shohei Koide¹^,⁴, Anthony Kossiakoff¹^,⁴, Sachdev S. Sidhu¹^,²

¹The Recombinant Antibody Network, ²The Banting and Best Department of Medical Research, University of Toronto, ³Antibiome Center, University of California, San Francisco at Mission Bay, ⁴Department of Biochemistry and Molecular Biology, The University of Chicago

Summary

A method is described with visual accompaniment for conducting scalable, high throughput selections from phage-displayed combinatorial synthetic antibody libraries against hundreds of antigens simultaneously. Using this parallel approach, we have isolated antibody fragments that exhibit high affinity and specificity for diverse antigens that are functional in standard immunoassays.

Explore More Videos

Synthetic Antibody Libraries

High throughput Selection

Antigen Characterization

Antibody Discovery

Chapters in this video

0:05

Title

2:17

Preparing Cells for Phage Selection

3:28

Library Pre-clearance and Antigen-phage Incubation

5:45

Phage Elution and Amplification

8:03

Characterization by Pooled ELISA

10:22

Anticipated Results and Troubleshooting

12:18

Conclusion

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