Evaluation of Solid Supports for Slide- and Well-Based Recombinant Antibody Microarrays.
Antibody microarrays have emerged as an important tool within proteomics, enabling multiplexed protein expression profiling in both health and disease. The design and performance of antibody microarrays and how they are processed are dependent on several factors, of which the interplay between the antibodies and the solid surfaces plays a central role. In this study, we have taken on the first comprehensive view and evaluated the overall impact of solid surfaces on the recombinant antibody microarray design. The results clearly demonstrated the importance of the surface-antibody interaction and showed the effect of the solid supports on the printing process, the array format of planar arrays (slide- and well-based), the assay performance (spot features, reproducibility, specificity and sensitivity) and assay processing (degree of automation). In the end, two high-end recombinant antibody microarray technology platforms were designed, based on slide-based (black polymer) and well-based (clear polymer) arrays, paving the way for future large-scale protein expression profiling efforts.
Department of Immunotechnology, Lund University, Medicon Village, Lund SE-22381, Sweden. anna.sandstrom_gerdtsson@immun.lth.se.|Clinical Cancer Research using Emerging Advanced Technologies for Health (CREATE), Lund University, Medicon Village, Lund SE-22381, Sweden. anna.sandstrom_gerdtsson@immun.lth.se.
Anna S
AS
Gerdtsson
Department of Immunotechnology, Lund University, Medicon Village, Lund SE-22381, Sweden. linda.dmellby@gmail.com.|Clinical Cancer Research using Emerging Advanced Technologies for Health (CREATE), Lund University, Medicon Village, Lund SE-22381, Sweden. linda.dmellby@gmail.com.
Linda
L
Dexlin-Mellby
Department of Immunotechnology, Lund University, Medicon Village, Lund SE-22381, Sweden. payam.delfani@immun.lth.se.|Clinical Cancer Research using Emerging Advanced Technologies for Health (CREATE), Lund University, Medicon Village, Lund SE-22381, Sweden. payam.delfani@immun.lth.se.
Payam
P
Delfani
Department of Immunotechnology, Lund University, Medicon Village, Lund SE-22381, Sweden. erica.berglund@immun.lth.se.
Erica
E
Berglund
Department of Immunotechnology, Lund University, Medicon Village, Lund SE-22381, Sweden. carl.borrebaeck@immun.lth.se.|Clinical Cancer Research using Emerging Advanced Technologies for Health (CREATE), Lund University, Medicon Village, Lund SE-22381, Sweden. carl.borrebaeck@immun.lth.se.
Carl A K
CA
Borrebaeck
Department of Immunotechnology, Lund University, Medicon Village, Lund SE-22381, Sweden. christer.wingren@immun.lth.se.|Clinical Cancer Research using Emerging Advanced Technologies for Health (CREATE), Lund University, Medicon Village, Lund SE-22381, Sweden. christer.wingren@immun.lth.se.
Christer
C
Wingren
affinity proteomics; antibody microarrays; array-in-well; microarray scanning; solid support; source plate