The analysis of interactions between molecules using surface plasmon resonance

Subject description

The method based on the surface plasmon resonance (SPR) is a modern biophysical analytical method for the analysis of interactions between molecules. Using the method we can analyze in the real time whether an interaction among selected (macro)molecules occurs and characterize the kinetics of interaction e.g. between proteins, proteins and membranes, proteins and nucleic acids, proteins and small molecules (bigger than 100 Da) and even among proteins and viruses or whole cells.

Within the theoretical part of the course the student learns:

(i) Basic parts and principle of operation of the SPR apparatus

(ii) Basic parameters of SPR analysis: Ka, Kd, ​​Rmax, KD, Chi2.

(iii) Set and chemistry of SPR chips used for the immobilization of molecules

(iv) How to design an experiment to analyse interactions between molecules

(v) Methods of immobilizing the ligand molecule on the selected SPR chip

(vi) Methods of injecting the analyte over the ligand and tracking the interactions in real-time

(vii) How to regenerate the chip surface and ensure repeatability of results

(viii) Data analysis – to determine whether ligand-analyte binding has occurred, analysis of binding kinetics of the interacting molecules.

The student practically designs and carries out the experiment and analyses the data of interaction between selected molecules.

The subject is taught in programs

Objectives and competences

Acquisition of knowledge about the ability to analyse interactions between (macro)molecules with an apparatus based on SPR.

Ability to conceive, perform protein-protein, protein-membrane, protein-DNA, protein-RNA, low-protein, cell lysate-protein or virus-protein interactions.

The student learns how to critically analyse and characterize properties of the interactions.

Teaching and learning methods

Consultations and assistance in the design, implementation and analysis of the SPR experiment of the interaction between selected molecules. In agreement with the candidate's mentor, it is possible to carry out a part of the research work of the student's doctoral dissertation in the SPR infrastructure center for measuring the molecular interactions.

Expected study results

The student performs an individual research task and »hands on« experiences the properties of the SPR apparatus. The student learns how to design and analyse the obtained measurements – the expected study result is a successful analysis of the selected interaction between molecules. The results of the analyses enable the student to design further experiments that would confirm the results of SPR analyses in vivo.

Basic sources and literature

Pregledni in izvirni članki iz področja.

Review and original articles from the field.


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