Automated panning and screening of antibodies from phage display libraries

Antti Tullila

Research output: Contribution to conferenceConference articleScientific

Abstract

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. We have developed and optimized 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 plate format. For screening, an enzyme-linked immunosorbent assay protocol suitable for a robotic station was developed. This system was first set up with protein and hapten molecules as model antigens to select antibodies from the VTT naive human single-chain antibody (scFv) library. Previously these antigens have been used successfully to select antibodies manually from the same library. The naïve library is modest in size (108), but yet panels of antigen specific antibody clones distinct by the amino acid sequences were achieved with this automated method. The selection and screening procedures were further developed and optimized for isolation of hapten specific antibodies recognizing solely the hapten conjugate and not the carrier protein. A multiplexed selection and screening was performed simultaneously for 12 hapten antigens differing by physical and chemical properties and size. The conjugation degree of each hapten to their carrier protein was validated by an MS analysis before the selection. Recombinant antibody fragments were successfully isolated against 8 of the 12 target antigens. All positive clones were produced efficiently in E. coli bacteria. The characterization of the binding properties of the best antibody clones for each hapten is in progress. In conclusion, the developed automated selection and screening method allows the efficient selection of antigen specific antibodies for multiple targets in parallel. It is also a powerful tool to screen concurrently various selection conditions for one antigen.
Original languageEnglish
Publication statusPublished - 2010
MoE publication typeNot Eligible
EventBIT Life Sciences' 2nd Annual International Congress of Antibody, ICA2010 - Beijing, China
Duration: 23 Mar 201025 Mar 2010
Conference number: 2

Conference

ConferenceBIT Life Sciences' 2nd Annual International Congress of Antibody, ICA2010
Abbreviated titleICA2010
CountryChina
CityBeijing
Period23/03/1025/03/10

Fingerprint

Bacteriophages
Libraries
Haptens
Antigens
Antibodies
Clone Cells
Robotics
Carrier Proteins
Single-Chain Antibodies
Immunoglobulin Fragments
Amino Acid Sequence
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Technology
Bacteria
Proteins

Cite this

Tullila, A. (2010). Automated panning and screening of antibodies from phage display libraries. Paper presented at BIT Life Sciences' 2nd Annual International Congress of Antibody, ICA2010, Beijing, China.
Tullila, Antti. / Automated panning and screening of antibodies from phage display libraries. Paper presented at BIT Life Sciences' 2nd Annual International Congress of Antibody, ICA2010, Beijing, China.
@conference{b819900d732742fda3db8cdf80f18bdd,
title = "Automated panning and screening of antibodies from phage display libraries",
abstract = "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. We have developed and optimized 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 plate format. For screening, an enzyme-linked immunosorbent assay protocol suitable for a robotic station was developed. This system was first set up with protein and hapten molecules as model antigens to select antibodies from the VTT naive human single-chain antibody (scFv) library. Previously these antigens have been used successfully to select antibodies manually from the same library. The na{\"i}ve library is modest in size (108), but yet panels of antigen specific antibody clones distinct by the amino acid sequences were achieved with this automated method. The selection and screening procedures were further developed and optimized for isolation of hapten specific antibodies recognizing solely the hapten conjugate and not the carrier protein. A multiplexed selection and screening was performed simultaneously for 12 hapten antigens differing by physical and chemical properties and size. The conjugation degree of each hapten to their carrier protein was validated by an MS analysis before the selection. Recombinant antibody fragments were successfully isolated against 8 of the 12 target antigens. All positive clones were produced efficiently in E. coli bacteria. The characterization of the binding properties of the best antibody clones for each hapten is in progress. In conclusion, the developed automated selection and screening method allows the efficient selection of antigen specific antibodies for multiple targets in parallel. It is also a powerful tool to screen concurrently various selection conditions for one antigen.",
author = "Antti Tullila",
year = "2010",
language = "English",
note = "BIT Life Sciences' 2nd Annual International Congress of Antibody, ICA2010, ICA2010 ; Conference date: 23-03-2010 Through 25-03-2010",

}

Tullila, A 2010, 'Automated panning and screening of antibodies from phage display libraries' Paper presented at BIT Life Sciences' 2nd Annual International Congress of Antibody, ICA2010, Beijing, China, 23/03/10 - 25/03/10, .

Automated panning and screening of antibodies from phage display libraries. / Tullila, Antti.

2010. Paper presented at BIT Life Sciences' 2nd Annual International Congress of Antibody, ICA2010, Beijing, China.

Research output: Contribution to conferenceConference articleScientific

TY - CONF

T1 - Automated panning and screening of antibodies from phage display libraries

AU - Tullila, Antti

PY - 2010

Y1 - 2010

N2 - 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. We have developed and optimized 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 plate format. For screening, an enzyme-linked immunosorbent assay protocol suitable for a robotic station was developed. This system was first set up with protein and hapten molecules as model antigens to select antibodies from the VTT naive human single-chain antibody (scFv) library. Previously these antigens have been used successfully to select antibodies manually from the same library. The naïve library is modest in size (108), but yet panels of antigen specific antibody clones distinct by the amino acid sequences were achieved with this automated method. The selection and screening procedures were further developed and optimized for isolation of hapten specific antibodies recognizing solely the hapten conjugate and not the carrier protein. A multiplexed selection and screening was performed simultaneously for 12 hapten antigens differing by physical and chemical properties and size. The conjugation degree of each hapten to their carrier protein was validated by an MS analysis before the selection. Recombinant antibody fragments were successfully isolated against 8 of the 12 target antigens. All positive clones were produced efficiently in E. coli bacteria. The characterization of the binding properties of the best antibody clones for each hapten is in progress. In conclusion, the developed automated selection and screening method allows the efficient selection of antigen specific antibodies for multiple targets in parallel. It is also a powerful tool to screen concurrently various selection conditions for one antigen.

AB - 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. We have developed and optimized 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 plate format. For screening, an enzyme-linked immunosorbent assay protocol suitable for a robotic station was developed. This system was first set up with protein and hapten molecules as model antigens to select antibodies from the VTT naive human single-chain antibody (scFv) library. Previously these antigens have been used successfully to select antibodies manually from the same library. The naïve library is modest in size (108), but yet panels of antigen specific antibody clones distinct by the amino acid sequences were achieved with this automated method. The selection and screening procedures were further developed and optimized for isolation of hapten specific antibodies recognizing solely the hapten conjugate and not the carrier protein. A multiplexed selection and screening was performed simultaneously for 12 hapten antigens differing by physical and chemical properties and size. The conjugation degree of each hapten to their carrier protein was validated by an MS analysis before the selection. Recombinant antibody fragments were successfully isolated against 8 of the 12 target antigens. All positive clones were produced efficiently in E. coli bacteria. The characterization of the binding properties of the best antibody clones for each hapten is in progress. In conclusion, the developed automated selection and screening method allows the efficient selection of antigen specific antibodies for multiple targets in parallel. It is also a powerful tool to screen concurrently various selection conditions for one antigen.

M3 - Conference article

ER -

Tullila A. Automated panning and screening of antibodies from phage display libraries. 2010. Paper presented at BIT Life Sciences' 2nd Annual International Congress of Antibody, ICA2010, Beijing, China.