Application Note

RSS

The ProteOn XPR36 protein interaction array system is a surface plasmon resonance (SPR) biosensor with a multichannel module and interaction array sensor chip for analysis of up to 36 protein interactions in a single injection step. The capability of the ProteOn XPR36 system to rapidly generate a 6 x 6 interaction array between six ligands and six analytes greatly increases the throughput, flexibility, and versatility of experimental design for a wide range of biomolecular interaction studies.

We describe here the rapid and detailed kinetic analyses of four antigen-antibody interactions. Briefly, the interaction of each antibody (ligand) with a concentration series of its respective antigen (analyte) was analyzed in a sequence of a single immobilization cycle followed by four analyte injection cycles. Four different antibodies and a nonreactive negative control protein were immobilized in five parallel ligand channels on a single ProteOn sensor chip; the sixth ligand channel was left blank for use as a reference channel. For each antibody, a concentration series of antigen samples (six samples spanning a wide concentration range) was then injected into the set of six parallel analyte channels orthogonal to the ligand channels. The ligand channels were then regenerated for analysis of the next antigen concentration series, until all four antigen-antibody interactions were analyzed.

Each of the four antigen injections produced a set of 36 sensorgrams. These contained data not only on the interaction of an antigen concentration series with its respective antibody, but also on its interaction with each of the three other antibodies, the negative control protein, and the reference channel. Thus, data sufficient to perform a detailed kinetic analysis of each antigen-antibody interaction — as well as a determination of any potential cross-reactivity among the antigens and antibodies — were obtained in a minimal number of injection steps.

The ProteOn XPR36 protein interaction array system is wellsuited for experimental designs in which a number of interacting protein pairs are to be analyzed and compared. Data can be rapidly obtained for protein interface characterization, affinity ranking, and epitope mapping of antibodies, as well as for selecting lead compounds in drug development.

The capability of the ProteOn XPR36 system to rapidly perform a detailed kinetic analysis of multiple protein-protein interactions in a 6 x 6 interaction array format is especially useful for antibody development and characterization.

Source: Bio-Rad Laboratories, Inc.

Download The Complete Application Note