| Chapter title |
Enzymatic Assembly for scFv Library Construction
|
|---|---|
| Chapter number | 3 |
| Book title |
Synthetic Antibodies
|
| Published in |
Methods in molecular biology, March 2017
|
| DOI | 10.1007/978-1-4939-6857-2_3 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6855-8, 978-1-4939-6857-2
|
| Authors |
Mieko Kato, Yoshiro Hanyu |
| Editors |
Thomas Tiller |
| Abstract |
Recombinant monoclonal antibodies can be established by displaying single-chain variable fragment (scFv) antibody libraries on phages and then biopanning against the target. For constructing superior scFv libraries, antibody light-chain variable region (VL) and heavy-chain variable region (VH) fragments must be assembled into scFvs without loss of diversity. A high-quality scFv library is a prerequisite for obtaining strong binders from the scFv library. However, the technical challenges associated with the construction of a diverse library have been the bottleneck in the establishment of recombinant antibodies through biopanning. Here, we describe a simple and efficient method for assembling VL and VH fragments through the concerted action of λ-exonuclease and Bst DNA polymerase. We successfully used this method to construct a diverse chicken scFv library. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 12 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 4 | 33% |
| Student > Bachelor | 2 | 17% |
| Professor > Associate Professor | 1 | 8% |
| Student > Master | 1 | 8% |
| Unknown | 4 | 33% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 4 | 33% |
| Veterinary Science and Veterinary Medicine | 1 | 8% |
| Agricultural and Biological Sciences | 1 | 8% |
| Medicine and Dentistry | 1 | 8% |
| Unknown | 5 | 42% |