Antibody Microarrays: Review of Technique

Antibody microarrays are used for profiling complex samples in large quantities at a time. The automation approach has been used to screen monoclonal antibody fragments for their roles and relation to each other. Unfortunately, there are limitations in the use of microarrays. To overcome the issue of microarrays only being able to analyze a few samples and lacking the ability of multiplexing capacity, the authors present the multiple spotting technique or MIST for short.

With MIST the authors attempt to illustrate automation extension by improving time, waste produced, and materials. The authors illustrate the use of MIST having the capability of screening thousands of monoclonal antibody fragments in a single chip. The authors expressed scFV on 384-well plates using 2YT medium with ampicillin, IPTG, glucose and glycerol in TGI cells. An ELISA for the 384 well plate was performed and the 384 well plate containing the cell cultures was used as a reference for spotting when they screened for binders on microarrays. Monoclonal anti-polyubiquitin scFv was prepared in a culture in order to compare the detection limit of the MIST technique to ELISA.

Their results produced two figures that denoted the MIST technique was actually a reasonable way to screen for recombinant antibody fragments. Figure 3 and 4 shows the results of the selection of a massive collection of recombinant antibody fragments. This part of the results show how the MIST technique can reduce the number of how many clones need to be screened and how the technique can be used to characterize fragments of different traits.

The authors conclude that their technique opens up different doors for different applications. Scvf applications on microarrays can be examined to override previous limitations and the clones that test positive can be characterized against many antigens by their technique, MIST. The article was written in a clear in a concise manner to which contributed the overall effectiveness; however, the conclusion was too brief for the amount of background information given that provided imbalance to the article and the future applications of the technique were not specified in any type of detail.

Agenendt, P.; Wilde, J.; Kijanka, G.; Baars, S.; Cahill, D.J.; Kreutzberger, J.; Lehrach, H.; Konthur, Z.; Glökler, J. Anal. Chem. 2004, 76, 2916-2921.