Antibody immobilization strategies

Peluso P., Wilson D.S., Do D., Tran H., Venkatasubbaiah M., Quincy D., Heidecker B., Poindexter K., Tolani N., Phelan M., Witte K., Jung L.S., Wagner P., Nock S., Optimizing antibody immobilization strategies for the construction of protein microarrays, Anal Biochem. 2003 312 113-124. 

Peluso P, Wilson DS, Do D, et al. (2003) Optimizing antibody immobilization strategies for the construction of protein microarrays. Anal Biochem 312, 113-24. 

Lee et al. utilized the self-assembled layer of thiol group-modified protein A for the oriented immobilization of antibodies [64], An increased binding capacity was further observed. As another illustrative instance, a protein A-based orientation-controlled immobilization strategy for antibodies was proposed for the fabrication of a QCM immunosensor using nanometer-sized gold particles and amine-terminated PPF [65], Moreover, in recent years, there has emerged another oriented immobilization... 

H. Wang, Y. Liu, Y. Yang, T. Deng, G.L. Shen, and R.Q. Yu, A protein A-based orientation-control-led immobilization strategy for antibodies using nanometer-sized gold particles and plasma-polymerized film. Anal. Biochem. 324, 219-226 (2004). 

A similar strategy was performed for the immunological determination of sulfonamides (SFM) in raw full cream, as well as in all varieties of ultra high temperature (UHT) milk, such as full cream (about 3.25% fat), semiskimmed (about 1.5-1.8% fat), and skimmed (0.1% fat), by using a class specific antisulfonamides (anti-SFM) antibodies immobilized on magnetic beads, and an SFM-HRP tracer for the electrochemical detection [62]. 

In recent years, a number of different types of chip surfaces have been investigated including glass or plastic slides, porous gel pad slides, filters, microwells, and multiple polymer surfaces [140-142]. However, the basis for antibody immobilization in these strategies has almost universally been nonspecific, that is, not leading to the oriented presentation of the antibody. The most frequently used method utilizes... 

FIGURE 7.7 Antibody orientation strategy for biochip development. Random immobilization by adsorption or covalent attachment leads to nonaccessible (A) or only partially accessible (B) antigen-binding domains. In (C) correct orientation of antibody will give maximum activity and this is achieved using a linker molecule (e.g., Protein A). 

---