Sekvensering av proteiner från spermatoforen hos Pieris napi genom automatiserad CE-fraktionering till en nyutvecklad AKD-belagd MALDI koncentreringsplatta, följt av trypsinklyvning, MS och MS/MS
Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
Sequencing of Pieris napi spermatophore proteins by automated CE-fractionation to a novel AKD coated MALDI concentration plate, followed by trypsin digestion, MS, and MS/MS (English)
A hydrophobic concentration plate for Matrix-Assisted Laser-Desorption Ionisation-Time-Of-Flight-Mass Spectrometry (MALDI-MS) has been developed and used for protein analysis.
The Alkyl Ketene Dimer (AKD) surface coating used gives a concentration effect that increases Signal to Noise ratio (S/N) of the measured peptides with more than a factor two compared to a steel plate, and surpasses a commercial concentration plate. The AKD MALDI concentration plate was used for off-line Capillary Electrophoresis (CE) to MALDI coupling in the analysis and partial sequencing of reproduction proteins in the spermatophore of recently mated females of Pieris napi (green-veined white butterfly). The extracted, concentrated and purified sample was analysed by MS and Tandem MS (MS/MS). Extracted and concentrated but not purified sample was then separated with CE and fractionated onto the AKD MALDI concentration plate for MS and MS/MS analysis. A second fractionated sample was trypsin digested before the same analysis. A database search of the sequences obtained by MS/MS gave no relevant homologies.
Place, publisher, year, edition, pages
2015. , 91 p.
Pieris napi, MALDI-TOF-MS/MS, AKD MALDI concentration plate, Automated CE-fractionation, Trypsin digestion
Engineering and Technology
IdentifiersURN: urn:nbn:se:kth:diva-172410OAI: oai:DiVA.org:kth-172410DiVA: diva2:847777