title: Phenylboronic acid polymer brush-enabled oriented and high density antibody immobilization for sensitive microarray immunoassay.
abstract: There still has a big challenge for low-abundance protein detection with antibody microarrays. In this work, phenylboronic acid (PBA) polymer brush-enabled oriented, high density, and covalent antibody immobilization was realized for sensitive antibody microarrays. PBA-enabled oriented antibody attachment via carbohydrate at Fc portion keeps antigen-binding sites fully expose to their corresponding antigens, resulting in higher antibody-antigen (Ab-Ag) binding efficiency. Sandwich immunoassay with rabbit IgG as model analyte was performed on poly(glycidyl methacrylate)-amino-phenylboronic acid-coated glass slide (PGMA-APBA-slide). One order improvement of LOD was achieved as compared with that on poly(glycidyl methacrylate) glass slide (PGMA-slide). The improvement is mainly attributed to PBA-assisted high density and oriented antibody immobilization. This work provides a versatile and effective strategy to develop high sensitive antibody microarrays for low-abundance protein analysis in various proteomic applications.
authors.affiliation: Institute for Clean Energy & Advanced Materials, Faculty of Materials & Energy, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, China; Chongqing Engineering Research Center for Rapid Diagnosis of Dread Diseases, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, China. Electronic address: ysliu@swu.edu.cn.
authors.forename: Yingshuai
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Institute for Clean Energy & Advanced Materials, Faculty of Materials & Energy, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, China; Chongqing Engineering Research Center for Rapid Diagnosis of Dread Diseases, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Institute for Clean Energy & Advanced Materials, Faculty of Materials & Energy, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, China; Chongqing Engineering Research Center for Rapid Diagnosis of Dread Diseases, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, China.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Institute for Clean Energy & Advanced Materials, Faculty of Materials & Energy, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, China; Chongqing Engineering Research Center for Rapid Diagnosis of Dread Diseases, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D000941:Antigens / Q000378:metabolism; D001897:Boronic Acids / Q000737:chemistry*; D005453:Fluorescence; D055767:Immobilized Proteins / Q000378:metabolism*; D007118:Immunoassay / Q000379:methods*; D011108:Polymers / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D011817:Rabbits; D059629:Signal-To-Noise Ratio; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D000906:Antibodies; D000941:Antigens; D001897:Boronic Acids; D055767:Immobilized Proteins; D011108:Polymers; C010686:benzeneboronic acid
keywords: Antibody microarray; Immunoassay; Oriented immobilization; Phenylboronic acid; Polymer brush