amino acids (PDZ-binding motif) are scrambled or the 2 or
3 last amino acids are omitted. We also recommend peptides to
be not shorter than 12–15 amino acids. In some cases peptide
as long as 30 amino acids were needed to observe interaction.
11. It is possible to observe “a bulk shift” at the beginning of the
injection. This can be due to differences in refractive index
between the running and the sample buffer.
12. It is important to determine a most appropriate regeneration
buffer (to choose the less aggressive regeneration conditions
allowing complete dissociation of the complex without
compromising the stability/properties of the ligand). A combination of high pH and high salt concentration is often used.
13. For lipids that have transition temperatures above room temperature it is recommended to place the extruder heating block
onto a warm plate.
14. To reduce the dead volume, pass a syringe with buffer through
the extruder.
15. For those lipids that have transition temperatures above room
temperature, allow the temperature of the lipid suspension to
equilibrate with the temperature of the heating block for at
least 5–10 min.
16. Lower immobilization levels are expected with mixtures of
negatively charged vesicles compared to liposomes composed
of zwitterion lipids.
17. When using single-cycle kinetics, one can proceed with up to
five analyte (increasing concentrations) injections.
18. L1 sensor chips can be reused several times. Yet more and more
liposomes will be required to reach 4000 RUs of
immobilization.
19. Analyte can be preincubated with a fixed concentration of
peptide before being perfused over immobilized liposomes.
Acknowledgments
Work in the laboratory of PZ is currently supported by the Fund for
Scientific Research–Flanders (FWO Grants G.0846.15 and
G0C5718N), the Agence Nationale de la Recherche (ANR-18CE13-0017, Project SynTEV) and receives funding from the
European Union’s Horizon 2020 research and innovation program
under the Marie Sklodowska-Curie grant agreement no. 747025.
86
Pascale Zimmermann and Antonio Luis Egea-Jimenez
3 last amino acids are omitted. We also recommend peptides to
be not shorter than 12–15 amino acids. In some cases peptide
as long as 30 amino acids were needed to observe interaction.
11. It is possible to observe “a bulk shift” at the beginning of the
injection. This can be due to differences in refractive index
between the running and the sample buffer.
12. It is important to determine a most appropriate regeneration
buffer (to choose the less aggressive regeneration conditions
allowing complete dissociation of the complex without
compromising the stability/properties of the ligand). A combination of high pH and high salt concentration is often used.
13. For lipids that have transition temperatures above room temperature it is recommended to place the extruder heating block
onto a warm plate.
14. To reduce the dead volume, pass a syringe with buffer through
the extruder.
15. For those lipids that have transition temperatures above room
temperature, allow the temperature of the lipid suspension to
equilibrate with the temperature of the heating block for at
least 5–10 min.
16. Lower immobilization levels are expected with mixtures of
negatively charged vesicles compared to liposomes composed
of zwitterion lipids.
17. When using single-cycle kinetics, one can proceed with up to
five analyte (increasing concentrations) injections.
18. L1 sensor chips can be reused several times. Yet more and more
liposomes will be required to reach 4000 RUs of
immobilization.
19. Analyte can be preincubated with a fixed concentration of
peptide before being perfused over immobilized liposomes.
Acknowledgments
Work in the laboratory of PZ is currently supported by the Fund for
Scientific Research–Flanders (FWO Grants G.0846.15 and
G0C5718N), the Agence Nationale de la Recherche (ANR-18CE13-0017, Project SynTEV) and receives funding from the
European Union’s Horizon 2020 research and innovation program
under the Marie Sklodowska-Curie grant agreement no. 747025.
86
Pascale Zimmermann and Antonio Luis Egea-Jimenez