2015
DOI: 10.3791/51492
|View full text |Cite
|
Sign up to set email alerts
|

Abstract: The demand for antibodies that fulfill the needs of both basic and clinical research applications is high and will dramatically increase in the future. However, it is apparent that traditional monoclonal technologies are not alone up to this task. This has led to the development of alternate methods to satisfy the demand for high quality and renewable affinity reagents to all accessible elements of the proteome. Toward this end, high throughput methods for conducting selections from phage-displayed synthetic a… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
2
2
1

Citation Types

0
23
0

Year Published

2015
2015
2023
2023

Publication Types

Select...
5
3
1

Relationship

2
7

Authors

Journals

citations
Cited by 22 publications
(23 citation statements)
references
References 25 publications
0
23
0
Order By: Relevance
“…Briefly E. coli cultures were grown in Terrific Broth with antibiotics to an OD 600 of 0.6–0.8 with 1 m m IPTG for 3–6 h at 37°C, then harvested by centrifugation. Cells were resuspended, lysed by sonication, and clarified prior to purification with GST Sepharose ( 24 ). As a second system we expressed the TFs as His-Avi-TEV N-terminal fusion driven from a T7 promoter using an E. coli in vitro transcription-translation system (IVTT) that was kindly provided by Sutro Biosciences ( 25 ) ( supplemental Fig.…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…Briefly E. coli cultures were grown in Terrific Broth with antibiotics to an OD 600 of 0.6–0.8 with 1 m m IPTG for 3–6 h at 37°C, then harvested by centrifugation. Cells were resuspended, lysed by sonication, and clarified prior to purification with GST Sepharose ( 24 ). As a second system we expressed the TFs as His-Avi-TEV N-terminal fusion driven from a T7 promoter using an E. coli in vitro transcription-translation system (IVTT) that was kindly provided by Sutro Biosciences ( 25 ) ( supplemental Fig.…”
Section: Methodsmentioning
confidence: 99%
“…All phage selections were done according to previously established protocols ( 24 , 26 ) with several modifications as outlined below. Briefly, Fab-phage selections were automated allowing for multi-parallel processing of biotinylated target antigens either generated in-house or obtained from collaborators.…”
Section: Methodsmentioning
confidence: 99%
“…The rationale is that when an organism is subjected to the appropriate environmental stress, it will generate mutations that convey the desired functional change (e.g., (29)). A variation of this approach is to build a library of protein mutants and screen or select for the desired function (e.g., (30,31)). Both approaches generate proteins with functional efficiencies similar to those of natural proteins, but they are limited by the number of mutants that can be sampled: a library of 10 10 variants only samples complete a.a. diversity at a maximum of seven to eight positions.…”
Section: Directed Evolutionmentioning
confidence: 99%
“…In vitro display technologies using yeast or phage are well-established approaches for generating high-affinity binding proteins from large naïve libraries. 8 In vitro selection can be done without the need for infected individuals and only requires the recombinant protein target. One of the recently developed modalities are small single domain antibodies derived from variable heavy homodimer (VHH) domains of antibodies from camels or llamas, often referred to as nanobodies, and are usually obtained by immunization and B-cell cloning.…”
Section: Introductionmentioning
confidence: 99%