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

Journal Article · Mon Jan 01 04:00:00 EST 2007 · Biochemistry and Molecular Biology Education

DOI:https://doi.org/10.1002/bmb.22· OSTI ID:912834

Kausaite, A ^[1]; van Dijk, M ^[2]; Castrop, J ^[2]; Ramanaviciene, A ^[1]; Baltrus, J P; Acaite, J ^[1]; Ramanavicius, A ^[1]

---

1. Vilnius Univ., Lithuania
2. Eco Chemie B.V., Utrecht, The Netherlands

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 biomolecular 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.

• Research Organization: Vilnius University, Lithuania; Eco Chemic B.V., Utrecht, The Netherlands; National Energy Technology Laboratory (NETL), Pittsburgh, PA, and Morgantown, WV
• Sponsoring Organization: Lithuanian State Science and Studies Foundation COST Program D31; USDOE - Office of Fossil Energy (FE)
• OSTI ID: 912834
• Report Number(s): DOE/NETL-IR-2007-063; Journal Serial ID: ISSN:: 1539-3429 : 1470-817
• Journal Information: Biochemistry and Molecular Biology Education, Journal Name: Biochemistry and Molecular Biology Education Journal Issue: 1 Vol. 35
• Publisher: John Wiley & Sons, Ltd.
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ALBUMINS
AMIDES
ANTIGENS
BIOCHEMISTRY
Biosensor
DETECTION
GOLD
IMMUNOLOGY
MOLECULAR BIOLOGY
MONITORING
MONITORS
PLASMONS
PROTEINS
RESONANCE
SPR
TRAINING
antibody
antigen
bioanalytical chemistry
education
surface plasmon resonance