Affinity-based Protein Immobilization and MALDI-TOF Analysis
Immobilization of the BTLA protein in a microcolumn and the affinity test were performed according to the procedure described in our previous studies [30 (link)]. The mass spectroscopy (MS) measurements were conducted using a MALDI-TOF/TOF 5800 (AB Sciex, Germany) instrument. α-Cyano-4-hydroxycinnamic acid (CHCA) was used as the matrix (10 mg/mL, Sigma-Aldrich, Saint Louis, MO, USA). The measurements were conducted in reflector positive mass mode with previous mass calibration with a commercial standard peptide mixture (The Peptide Mass Standards Kit for Calibration of AB SCIEX MALDI-TOF™ Instruments).
Spodzieja M., Kuncewicz K., Sieradzan A., Karczyńska A., Iwaszkiewicz J., Cesson V., Węgrzyn K., Zhukov I., Maszota-Zieleniak M., Michielin O., Speiser D.E., Zoete V., Derré L, & Rodziewicz-Motowidło S. (2020). Disulfide-Linked Peptides for Blocking BTLA/HVEM Binding. International Journal of Molecular Sciences, 21(2), 636.
Btla Hydroxycinnamic acidImmobilization Maldi Mass spectroscopy Peptide Protein
Corresponding Organization : University Hospital of Lausanne
Other organizations :
SIB Swiss Institute of Bioinformatics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Adam Mickiewicz University in Poznań, Ludwig Cancer Research, University of Lausanne
Immobilization of the BTLA protein in a microcolumn
dependent variables
Affinity test
control variables
Mass spectroscopy (MS) measurements conducted using a MALDI-TOF/TOF 5800 (AB Sciex, Germany) instrument
α-Cyano-4-hydroxycinnamic acid (CHCA) used as the matrix (10 mg/mL, Sigma-Aldrich, Saint Louis, MO, USA)
Measurements conducted in reflector positive mass mode
Mass calibration with a commercial standard peptide mixture (The Peptide Mass Standards Kit for Calibration of AB SCIEX MALDI-TOF™ Instruments)
controls
Positive control: Commercial standard peptide mixture used for mass calibration
Negative control: Not explicitly mentioned
Annotations
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