In the Biomolecular Interaction Laboratory we characterize the interaction of biomolecules using Biacore-3000 instrument, MicroCal iTC200, Monolith NT.115 and NT.LabelFree equipments. In addition, we have a Prometheus NT.48 device for the analysis of protein stability and heat denaturation. These devices are suitable for quantitative determination of binding constants as well as kinetic and thermodynamic characteristics.
Equipments
Biacore 3000
- Surface Plasmon Resononance based binding technique
- interaction analysis with immobilized ligand
- integrated microfluidic system
- different sensor chips available
- the kinetics of interaction (ka és kd)
- the affinity of interactions (KA=ka/kd or KD=kd/ka)
- thermodynamics (∆G, ∆H, ∆S)
- specificity of interactions (identification of interacting regions)
- identification of interacting partners from lysates
ITC MicroCal200
- ligand in syringe (Injected into sample cell)
- protein target in sample cell
- heat release or intake of interaction is measured
- hastelloy cell
- cell volume: 200 μL
- sensitive
- multiple feedback modes
- affinity KD from the millimolar (mM) to nanomolar (nM) range
- number of binding sites (N)
- thermodynamics: binding enthalpy (ΔH) and entropy (ΔS)
Monolith NT.115
- low sample volume (capillary volume ~6-8 μL)
- quantifying the interactions of purified proteins or in complex bioliquids
- Amine-reactive, cysteine-reactive or His-tag specific labeling using NanoTemper Protein Labeling Kits
- affinity (KD)
Monolith NT.LabelFree
- low sample volume (capillary volume ~6-8 μL)
- without protein labelling (one interacting partner contains intrinsic fluorofores)
- affinity (KD)
Prometheus NT.48
- protein heat stability screening (melting point TM)
- from 15°C to 95°C
- heating rate: 0.1 to 7°C/min
- measure 48 capillaries „on-the-fly“ with in 3 seconds
- low sample volume (capillary volume ~8-10 μL)
- High-Resolution Biophysical Characterization
- multi-domain unfolding transitions
- antibodies + antibody drug conjugates
- quality control
- forced degradation
- long-term stability of biologicals
Contact: erdodi@med.unideb.hu
The following charges are applied for the services
|
Work |
Unit |
Price (HUF) |
|
Biacore-3000 |
||
|
Equipment usage time |
hour |
10 000 |
|
Immobilization |
1 chip/4 surfaces |
15 000 |
|
Data evaluation |
1 concentration series |
18 000 |
|
|
(min. 3, max 7 conc.) |
|
|
Evaulation time |
hour |
5 000 |
|
MicroCal ITC200 |
||
|
Equipment usage time |
hour |
10 000 |
|
Data evaluation |
1 ligand titration with 1 interacting partner |
5 000 |
|
Monolith NT.115 and NT.LabelFree |
||
|
Equipment usage time |
hour |
10 000 |
|
Standard Capillaries |
16 pieces per measurement |
2 600 |
|
Premium Capillaries |
16 pieces per measurement |
11 500 |
|
Capillaries Monolith NT.LabelFree |
16 pieces per measurement |
7 800 |
|
Premium Capillaries Monolith NT.LabelFree |
16 pieces per measurement |
13 500 |
|
Protein labelling |
per protein |
35 000 |
|
Data evaluation |
hour |
5 000 |
|
Prometheus NT.48 |
||
|
Equipment usage time |
hour |
10 000 |
|
Standard Capillaries |
3 pieces of per sample |
750 |
|
High Sensitivity Capillaries |
3 pieces of per sample |
2 700 |
|
Data evaluation |
hour |
5 000 |