High-throughput affinity ranking of antibodies using surface plasmon resonance microarrays

Wassaf, Dina; Kuang, Guannan; Kopacz, Kris; Wu, Qilong; Nguyen, Que Thanh Thanh; Toews, Mark; Cosic, Janja; Jacques, Judith; Wiltshire, Steve; Lambert, Jérémy; Pazmany, Csaba; Hogan, Sean A.; Ladner, Robert C.; Nixon, Andrew E.; Sexton, Daniel J.

doi:10.1016/j.ab.2006.01.043

Cited by 123 publications

(81 citation statements)

References 36 publications

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“…competitive eLISA tests are often very unreliable compared to surface plasmon resonance (SpR) data. 28 SpR measurements, on the other hand, require the antibody to be purified, although there is one very recent report on affinity ranking of scfv fragments from crude lysates. 29 Two recent examples of high-throughput purification methods for antibodies exist.…”

Section: Discussionmentioning

confidence: 99%

Automated Panning and Screening Procedure on Microplates for Antibody Generation from Phage Display Libraries

Turunen¹,

Takkinen²,

Söderlund³

et al. 2009

SLAS Discovery

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Antibody phage display technology is well established and widely used for selecting specific antibodies against desired targets. Using conventional manual methods, it is laborious to perform multiple selections with different antigens simultaneously. furthermore, manual screening of the positive clones requires much effort. The authors describe optimized and automated procedures of these processes using a magnetic bead processor for the selection and a robotic station for the screening step. Both steps are performed in a 96-well microplate format. In addition, adopting the antibody phage display technology to automated platform polyethylene glycol precipitation of the enriched phage pool was unnecessary. for screening, an enzyme-linked immunosorbent assay protocol suitable for a robotic station was developed. This system was set up using human γ-globulin as a model antigen to select antibodies from a VTT naive human single-chain antibody (scfv) library. In total, 161 γ-globulin-selected clones were screened, and according to fingerprinting analysis, 9 of the 13 analyzed clones were different. The system was further tested using testosterone bovine serum albumin (BSA) and β-estradiol-BSA as antigens with the same library. In total, 1536 clones were screened from 4 rounds of selection with both antigens, and 29 different testosterone-BSA and 23 β-estradiol-BSA binding clones were found and verified by sequencing. This automated antibody phage display procedure increases the throughput of generating wide panels of target-binding antibody candidates and allows the selection and screening of antibodies against several different targets in parallel with high efficiency. (Journal of Biomolecular Screening 2009:282-293)

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Section: Discussionmentioning

confidence: 99%

Automated Panning and Screening Procedure on Microplates for Antibody Generation from Phage Display Libraries

Turunen¹,

Takkinen²,

Söderlund³

et al. 2009

SLAS Discovery

View full text Add to dashboard Cite

show abstract

“…Function-based primary screening of clones is often performed in an ELISA format, but because binding measurements in microwell plates require relatively large volumes of material, many wash steps, and are not particularly sensitive, sequencebased primary screening has predominated. Others have recognized the need for more efficient functional assessment, and alternative approaches such as SPR (11) have been employed for the secondary characterization of expressed single chain antibodies derived from phage screens, but these still employ an ELISA-based primary screening step prior to characterization.…”

Section: Discussionmentioning

confidence: 99%

High‐throughput screening and characterization of clones selected from phage display libraries

Yang

Nolan

2007

Cytometry Pt A

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Clones from phage display libraries are generally selected by a number of rounds of panning and regrowth, followed by primary screening to identify hits and secondary characterization to identify clones with optimal affinity and specificity. Because functional screening for binding or other activity can be material-, time-, and labor-intensive, sequencing is often used to identify the emergence of a consensus sequence prior functional characterization. However, the consensus sequence is not always the optimal one because factors such as phage growth rates, nonspecific binding, and other selection pressures can bias the selection process. To improve function-based phage display library screening and characterization, we developed a multiplexed approach employing optically-encoded microsphere arrays and flow cytometry. We show that capture of phage from crude culture supernatants enables the efficient screening of binding activity and the evaluation of binding avidity. The approach uses small volumes and a homogeneous no-wash format that minimizes reagent consumption and sample handling. The use of opticallyencoded microspheres allows many phage to be screened simultaneously, greatly increasing throughput. This approach is flexible, supporting primary and secondary screening for a range of functional assays, and scalable, potentially supporting the screening of thousands to hundreds of thousands of clones per hour. ' 2007

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“…The application of an antibody microarray system in which SPR technology is used for signal detection can dramatically increase the throughput compared to Biacore instruments and was successfully used to screen hybridoma supernatants. 28 There are a number of limitations to SPR and other kineticbased technologies (KinExA). For example, assay configuration and design are extremely important to avoid misinterpretation of reaction data.…”

Section: Discussionmentioning

confidence: 99%

High-Throughput Kinetic Screening of Hybridomas to Identify High-Affinity Antibodies Using Bio-Layer Interferometry

et al. 2015

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High-throughput affinity ranking of antibodies using surface plasmon resonance microarrays

Cited by 123 publications

References 36 publications

Automated Panning and Screening Procedure on Microplates for Antibody Generation from Phage Display Libraries

Automated Panning and Screening Procedure on Microplates for Antibody Generation from Phage Display Libraries

High‐throughput screening and characterization of clones selected from phage display libraries

High-Throughput Kinetic Screening of Hybridomas to Identify High-Affinity Antibodies Using Bio-Layer Interferometry

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