Surface plasmon resonance label-free monitoring of antibody antigen interactions in real time

被引:59
作者
Kausaite, Asta
van Dijk, Martijn
Castrop, Jan
Ramanaviciene, Almira
Baltrus, John P.
Acaite, Juzefa
Ramanavicius, Arunas
机构
[1] Vilnius State Univ, Inst Immunol, Lab Immunoanal & Nanotechnol, LT-08409 Vilnius 21, Lithuania
[2] Vilnius State Univ, Dept Analyt & Environm Chem, LT-03225 Vilnius 6, Lithuania
[3] Eco Chem BV, NL-3526 KM Utrecht, Netherlands
[4] US DOE, Natl Energy Technol Lab, Pittsburgh, PA 15236 USA
关键词
biosensor; bioanalytical chemistry; surface plasmon resonance; SPR; antibody; antigen; education;
D O I
10.1002/bmb.22
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Detection of biologically active compounds is one of the most important topics in molecular biology and biochemistry. One of the most promising detection methods is based on the application of surface plasmon resonance for label-free detection of biologically active compounds. This method allows one to monitor binding events in real time without labeling. The system can therefore be used to determine both affinity and rate constants for interactions between various types of molecules. Here, we describe the application of a surface plasmon resonance biosensor for label-free investigation of the interaction between an immobilized antigen bovine serum albumin (BSA) and antibody rabbit anti-cow albumin IgG1 (anti-BSA). The formation of a self-assembled monolayer (SAM) over a gold surface is introduced into this laboratory training protocol as an effective immobilization method, which is very promising in biosensing systems based on detection of affinity interactions. In the next step, covalent attachment via artificially formed amide bonds is applied for the immobilization of proteins on the formed SAM surface. These experiments provide suitable experience for postgraduate students to help them understand immobilization of biologically active materials via SAMs, fundamentals of surface plasmon resonance biosensor applications, and determination of non-covalent biornolecular interactions. The experiment is designed for master and/or Ph.D. students. In some particular cases, this protocol might be adoptable for bachelor students that already have completed an extended biochemistry program that included a background in immunology.
引用
收藏
页码:57 / 63
页数:7
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