Chapter title |
Yeast display of engineered antibody domains.
|
---|---|
Chapter number | 5 |
Book title |
Therapeutic Proteins
|
Published in |
Methods in molecular biology, June 2012
|
DOI | 10.1007/978-1-61779-921-1_5 |
Pubmed ID | |
Book ISBNs |
978-1-61779-920-4, 978-1-61779-921-1
|
Authors |
Zhao Q, Zhu Z, Dimitrov DS, Qi Zhao, Zhongyu Zhu, Dimiter S. Dimitrov, Zhao, Qi, Zhu, Zhongyu, Dimitrov, Dimiter S. |
Abstract |
Yeast display is an efficient technology for selection of antibodies and other proteins with high affinity and thermal stability. Here, we describe a method for affinity maturation of engineered antibody domains (eAds) using yeast display. EAd yeast libraries of relatively large size (∼10⁹) were generated and subjected to alternating rounds of magnetic-activated cell sorting (MACS), fluorescent-activated cell sorting (FACS), and random mutagenesis. The highest affinity clones from the final round of maturation were identified and analyzed. We discuss extensively each key step, and provide detailed protocols and helpful notes. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 21 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 5 | 24% |
Researcher | 5 | 24% |
Student > Bachelor | 2 | 10% |
Professor | 1 | 5% |
Unspecified | 1 | 5% |
Other | 2 | 10% |
Unknown | 5 | 24% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 5 | 24% |
Biochemistry, Genetics and Molecular Biology | 4 | 19% |
Unspecified | 1 | 5% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 5% |
Chemical Engineering | 1 | 5% |
Other | 3 | 14% |
Unknown | 6 | 29% |