Work

Investigation of Non-Nucleophilic Additives for the Reduction of Morphological Anomalies in Protein Arrays

Year: 2008

Type: article

Abstract: Uniform spot morphology is of critical importance in the fabrication and successful use of protein arrays, and solution additives are often needed to ensure good spot quality. Whereas hydroxyl-bearing molecules such as glycerol have found wide use, in our experience these reduce the efficiency of probe immobilization (particularly in the context of aldehyde-terminated surfaces). Here, we report a series of non-nucleophilic molecules that can be used as additives to improve spot homogeneity in protein arrays. Arrayed imaging reflectometry, a label-free optical biosensing technique, has been used along with spectroscopic ellipsometry to test the spot homogeneity, antibody immobilization efficiency, and activity of antihuman IgG arrays prepared with these non-nucleophilic additives on glutaraldehyde surfaces. It has been determined that 0.1% v/v 12-crown-4 performs optimally in MPBS buffer.

Source: Langmuir

DOI: 10.1021/la801712m

Authors: Charles R. Mace, Amrita R. Yadav, Benjamin L. Miller

Institutions: University of Rochester Medicine, University of Rochester

Language: English

Topic: Advanced Biosensing Techniques and Applications

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: U.S. Public Health Service, National Institute of Allergy and Infectious Diseases, National Center for Research Resources

Awards: G1960547943, G492466764, The University of Rochester Clinical and Translational Science Institute