Preparation of substrates for microarray protein chips with different ending functional groups

J Immunol Methods. 2022 Mar:502:113218. doi: 10.1016/j.jim.2022.113218. Epub 2022 Jan 10.

Abstract

Protein microarray chips are composed of three components, these are pre-treatment substrates, surface chemical modification, and immobilizing protein on substrate surfaces. In this study, self-assembly monolayers are used for surface chemical modification. Using this method, silanization on a glass and silicon chip is achieved, forming the terminal group substrates. Modification of the substrate surface to provide COOH and NH2 terminal functional groups provides a mechanism to proteins to immobilize on the substrate surface. To observe immobilized proteins on the substrate surface, they are first labeled with Cy5 fluorescent dye before analysis using a GenePix 4000B Microarray Scanner. The scanner induces fluorescence in the labelling dye and the resulting light is analyzed to provide information concerning both the quantity of immobilized protein, and the orientation of attachment. The antigen of the HSV-1 virus, a common human virus, was used in this study, performing an antigen-antibody analysis to determine the efficacy of the method under test for clinical diagnosis.

Keywords: Cy5-protein; Microarray protein substrates; Self-assembly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens
  • Fluorescent Dyes
  • Humans
  • Protein Array Analysis* / methods
  • Proteins*
  • Silicon
  • Surface Properties

Substances

  • Antigens
  • Fluorescent Dyes
  • Proteins
  • Silicon