Download - Sheldon Biotechnology Centre Mass Spectroscopy · Sheldon Biotechnology Centre The Sheldon Biotechnology Centre selVes as the core facility for biotechnology research at McGill University.

Transcript
Page 1: Sheldon Biotechnology Centre Mass Spectroscopy · Sheldon Biotechnology Centre The Sheldon Biotechnology Centre selVes as the core facility for biotechnology research at McGill University.

l

Sheldon Biotechnology CentreThe Sheldon Biotechnology CentreselVes as the core facility forbiotechnology research at McGillUniversity. We have state-of-the-arttechnical support facilities andoffer reliable low cost selVice toinvestigators.SelVices available at the SheldonBiotechnology Centre are oligo-nucleotide synthesis/purification,peptide synthesis, protein/peptidesequencing, protein/peptide aminoacid analysis and mass spectroscopy.

We are dedicated to remaining inthe forefront of recombinant DNAmethodology, protein sequencinganalysis and protein separations.This objective will selVe investiga-tors undertaking new studies andin the transfer of informationbetween various scientific disciplines.

Collaborative ProgramThe Sheldon Biotechnology Centrerepresents McGill University and itsaffiliated hospitals and researchinstitutes to the larger communityon matters relating to biotechno-logy. We recognize the need toform strong relationships withcolleagues in industry. Our Biotech-nology Collaborative Programfacilitates and coordinates thedevelopment of joint venturesbetween the business communityand McGill researchers working inthe field of biotechnology.

Victor L. Cape Lecture SeriesThe focus of this series is onbiotechnology. The program pro-vides a forum for industry, govern-

ment and university to meet anddiscuss mutual interests in the field.Recent speakers have includedProfessor Luc Montagnier, PasteurInstitute whose topic was"Biotechnology in AIDS Diagnosisand Prevention"; Dr. Webster K.Cavenee, Director, Ludwig Institutefor Cancer Research whose topicwas "Molecular Genetics of HumanCancer" and Dr. Leroy E. Hood,Director, Science and TechnologyCenter for Biotechnology whosetopic was "Biotechnology, theGenome Initiative and Medicine inthe Twenty-first Century". We striveto maintain a high standard ofinternationally recognized speakers.

Teaching Programin BiotechnologyThe Minor Program in Biotech-nology is administered for theFaculties of Engineering, Medicine,Agriculture and Science by theDepartment of Biology. ThisProgram allows students to select apackage of courses that willbroaden their perspective in areasimpacting on biotechnology.

Biotechnology 505B is a keycomponent of the Minor Programin Biotechnology and an importantcourse for graduate studentsseeking to expand their knowledgein biotechnology. This multidisci-plinary course is taught by out-standing McGill professors andresearchers from industry. Eachlecturer presents the significance ofhis or her own research to the fieldof biotechnology.

Mass SpectroscopyApplication of mass spectroscopy(MS) to protein characterization isa relatively new field. Severalapplications have been realizedand are routinely available, how-ever many new applications are yetto be realized.

Mass spectroscopy is readily appliedto quality control of syntheticpeptides, characterization of wholeprotein digests, identification ofpost-translational modifications, etc.

The Sheldon Biotechnology Centrehas access to two mass spectrom-eters which employ different'

ionization technologies: fast atombombardment (FAB)and electro-spray (ES) ionizations. FAB-MSisuseful in the low mass range andmay provide fragmentation infor-mation. ES-MSis a milder ioniza-tion technique and allows largermolecules (up to 100,000 amu) tobe characterized. Sample analysisand data evaluation are providedfor both technologies.

Emphasis is placed on assessingthe protein related requirementand providing guidance for appro-priate mass spectral analysis.

OrderingAll work is performed upon receiptof a valid purchase order. To obtainorder forms and price lists, pleasecontact the Sheldon BiotechnologyCentre by telephone (514) 398-3998or by fax (514) 398-8069.

Telephone orders can be placedMonday through Friday duringregular business hours. Fax orderscan be placed 24 hours a day.The staff of the SheldonBiotechnology Centre respectsthe confidentiality of researchinformation and projects.

For more information:SHELDON BIOTECHNOLOGY CENTREMcGill University3773 University StreetMontreal, Quebec H3A 2B4CanadaTel.: (514) 398-3998Fax: (514) 398-8069or return the attached card,indicating your area of interest.

Acknowledgements

The Sheldon Biotechnology Centreacknowledges the following fortheir support: Dr. HuntingdonSheldon; Pharmacia Canada Inc.;Hewlett-Packard Canada; the Deansof the Faculties of Science, Medicine,Agriculture and Graduate Studiesand Research of McGill University;the Research Institutes of theMontreal Children's Hospital andthe Royal Victoria Hospital. TheSheldon Biotechnology Centre hasalso received MRC/NSERCEquip-ment and Salary Support Grants.

Page 2: Sheldon Biotechnology Centre Mass Spectroscopy · Sheldon Biotechnology Centre The Sheldon Biotechnology Centre selVes as the core facility for biotechnology research at McGill University.

,-

Protein/Peptide SequencingI'rotdn/pt:ptidt: st:quencing assists At the Sheldon Biotechnology Centreinvestigators with separation we use Porton Protein/Peptidemethods and physical analysis of MicroSequencers with an integratedproteins. Sample purity and quan- customized microbore HPLC totitation are critical parameters optimize sequence. Direct on-linewhich strongly influence data analysis allows us to sequenceassessment and reliability particu- picomole amounts of protein withlady at the picomole level. exceptional results.

This facility serves as the resource Our protein/peptide sequencingfor peptide and protein isolation facility provides direct sequencestrategies using classical as well as analysis of isolated peptide andhigh performance liquid chromatog- protein fragments and interpretationraphy procedures. of the amino acid sequence data.

We provide advice in the prepara- High quality reliable data routinelytion of samples for sequencing and obtained by the Porton Gas Phasein methods of purification to those Sequencer is evaluated by comput-unfamiliar with the techniques of erized techniques and by an expe-protein chemistry. We strongly rienced sequencer. Qualification ofrecommend consultation with the the results with respect to sampleFacility Manager regarding compat- purity and quantitation is empha-ibility of your protein/peptide sized when reported to the client.purification protocol with high sen-sitivity protein/peptide sequencing.

Oligonucleotide Synthesis/PurificationSynthetic oligonucleotides areessential reagents to the molecularbiologist and are used in a widevariety of recombinant DNA manip-ulations, including DNA sequenc-ing, PCR amplification, site-directedmutagenesis, etc.

The Sheldon Biotechnology Centrehas capabilities for high throughputand flexibility. Using the GeneAssemble~ Plus, a 2 column DNAsynthesizer, we can provide youwith oligonucleotides of lengths toover 100 bases that include regularor modified bases such as inosineor methylated derivatives. On-linemonitoring during automatedsynthesis and purification by FastProtein Liquid Chromatography

Peptide SynthesisA very broad range of applicationsrequire synthetic peptides. Peptidesynthesis can be time consumingwithout specialized scientists andproper equipment.

At the Sheldon BiotechnologyCentre we have state of the arttechnology and peptide chemiststhat guarantee the synthesis. Solidphase peptide synthesis is carriedout on the Biolynx™ PeptideSynthesizer which uses the Fmocpolyamide peptide synthesismethodology. Up to 3 columns inseries can be used for productionof peptide analogues with yields ofapproximately 0.2 mmole/column.We can synthesize up to 100 mg ofpeptide for all types of applications.

The purity of the cleaved and

(FPLC) give you complete assur-ance of successful synthesis, highyield and purity.

Oligonucleotides are cleaved anddeprotected at 550C for 16 hoursin concentrated ammoniumhydroxide. Purification by FPLCcan be completed within 24 to 48hours following deprotection.

The Sheldon Biotechnology Centreis the source for affordable syn-thetic oligonucleotides resulting inconsiderable savings to investigators.All oligonucleotides are guaran-teed. Should there be a problem,we will re-synthesize and/or re-purify without charge.

deprotected peptide is assessedusing the HPLC DfB System Twowhich is equipped with a RapidSpectral Detector. Depending onthe sequence, the purity of thecrude peptide varies between 70%and 95%. If your applicationrequires greater purity, we offercomplete purification and charac-terization services. We have capa-bilities for carrying out qualitycontrol at the level of amino acidanalysis, mass spectroscopy andlO-peptide sequencing, dependingon your needs.

Please contact the Facility Managerconcerning specific additionalrequirements such as peptidecouplings or incorporation ofmodified amino acid residues.

Protein/Peptide Amino Acid AnalysisAmino acid analysis is one of themost exacting application areas ofliquid chromatography. Resultshave to be both accurate andreproducible.

At the Sheldon BiotechnologyCentre, we use the Alpha PlusAmino Acid Analyzer. With thissystem, the amino acids of the mostcomplex protein hydrolysates orphysiological fluids can be analyzed.

If your demand is perfection in thechromatography of protein/peptidehydrolysates you will be impressedwith the excellent resolution thesulphonated polystyrene resinprovides. This is an important

advantage where samples are notpure proteins but contain manyother components. Hydrolysistime-course profiles and/or pre-hydrolysis chemical modificationsare available for exacting quantita-tion of residues modified, destroyedor slowly released during routinehydrolysis conditions.

For complex biological fluids, weoffer baseline separation betweenvirtually all 40 of the amino acidsand their derivatives which canoccur in physiological fluids. Thisis ideal for complex samples suchas urine, plant extracts and fermen-tation media.