Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 282-293 |
| Number of pages | 12 |
| Journal | Journal of Biomolecular Screening |
| Volume | 14 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2009 |
| MoE publication type | A1 Journal article-refereed |
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Keywords
- antibody phage display
- automation
- high throughput
- magnetic beads
- robotic station
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Automated panning and screening procedure on microplates for antibody generation from phage display libraries. / Turunen, Laura (Corresponding Author); Takkinen, Kristiina; Söderlund, Hans; Pulli, Timo.
In: Journal of Biomolecular Screening, Vol. 14, No. 3, 2009, p. 282-293.Research output: Contribution to journal › Article › Scientific › peer-review
TY - JOUR
T1 - Automated panning and screening procedure on microplates for antibody generation from phage display libraries
AU - Turunen, Laura
AU - Takkinen, Kristiina
AU - Söderlund, Hans
AU - Pulli, Timo
PY - 2009
Y1 - 2009
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. 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.
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. 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.
KW - antibody phage display
KW - automation
KW - high throughput
KW - magnetic beads
KW - robotic station
U2 - 10.1177/1087057108330113
DO - 10.1177/1087057108330113
M3 - Article
VL - 14
SP - 282
EP - 293
JO - Journal of Biomolecular Screening
JF - Journal of Biomolecular Screening
SN - 1087-0571
IS - 3
ER -