Protein Purification Brochure

ProteinPurification: principles and PracticeThe State University of New Jersey - Rutgers Campus at New BrunswickA Five and One-Half DayHands-OnLaboratory CourseThe Center for Researchand Education in Bioluminescenceand Biotechnology(CREBB)Presents:Participants will learn skills to effectivelydesign separation and purification strate-gies for downstream processing. Thiscourse uses GFP (Green-FluorescentProtein), a novel marker for gene expres-sion, as the source material. While this is ageneral course applicable to all proteins,we use GFP as a highly effective visualteaching tool.Limited EnrollmentJune 1 - June 6, 2008July 13 - July 18, 2008July 27 - August 1, 2008January 11 - January 16, 2009March 15 - March 20, 2009Summer 2009 TBA www.rci.rutgers.edu/~crebb/protein.htmlDr. William W. Ward,Associate Professor ofBiochemistrySchool of Environmentaland Biological Sciences,Rutgers Universityand Director of C.R.E.B.BInstructorsFaculty CoordinatorThe Center for Research and Education inBioluminescence and Biotechnology(CREBB)The Center offers a series of continuing educationworkshops each year featuring nationally renownedpresenters. The Center for Research and Educationin Bioluminescence and Biotechnology (CREBB) is acomponent of Rutgers University, School of Environ-mental and Biological Sciences. The CREBB missionis to perform basic research on bioluminescence andto utilize bioluminescence (especially the Green Fluo-rescent Protein) as a tool to educate the scientific andindustrial communities in the field of biotechnology.Dr. Ward is a noted teacher and seminarleader with more than 20 years of experi-ence with adult professional audiences. He created "Protein Purifi-cation" in 1989 and the course has been running continuously ever since,reaching an estimated audience of over 1,200 scientists from across theglobe. Dr. Ward specializes in the chemical and physical properties of thegreen-fluorescent protein and other proteins involved in marine biolumi-nescence. He is published in 112 books, book chapters, journal articlesand abstracts. Dr. Ward has co-authored (with Catherine Thomson, Ph.D.)"A Guide to Green-Fluorescent Protein: Applications in Cell Biology andDrug Discovery" for D&MD Publications.This resource-oriented practi-cal guide presents an overview of the most important features and tech-nological applications of Green-Fluorescent Protein (GFP) and its vari-ant forms, and illustrates how GFP is currently being used in thebiopharmaceutical industry. He has also authored "Biochemical and PhysicalProperties of Green-Fluorescent Proteins", a chapter in "Green-Fluores-cent Protein: Properties, Applications, and Protocols, 2nd Edition",Chalfie and Kain (eds.), Wiley-Liss, Inc. 2006.Additional InstructorsDr. Gavin Swiatek, Biochemistry Instructor,School of Environmentaland Biological Sciences, Rutgers UniversityDr. Caren Villano, Biochemistry Instructor,School of Environmentaland Biological Sciences, Rutgers UniversityDr. Richard Ludescher, Professor of Food Science, Cook College,Rutgers UniversityMr. Robert Muldowney, Associate Director, Structural BiologyComputational Laboratory, Cook College, Rutgers UniversityDr. Olubayi Olubayi, Asst. Professor, Middlesex Community CollegeMore than 1,200 scientists from around the world havestrongly recommended this intensive course as an oppor-tunity to develop protein research and analytical skills in a re-treat setting. Participants work hard, identify and solve prob-lems in the lab and enjoy camaraderie and good food and drinkwith colleagues.This five-day laboratory course covers a wide variety ofconventional methods for protein isolation, purification, andcharacterization. The course format integrates hands-on labora-tory exercises with classroom lectures, demonstrations, studybreaks, and short take-home assignments.A special feature of the course is that all laboratory work will be performed on the same starting sample (AequoreaGFP or recombinant GFP), which will be purified from an exceed-ingly crude form (starting with tissue or bacterial cell extraction)to near homogeneity as judged by high performance liquid chro-matography (HPLC), SDS gel electrophoresis, isoelectric focus-ing, and Western blotting. This feature provides a continuity ofpurpose, integrating dozens of preparative and analytical pro-tein techniques in a way that few competing courses can match.A problem-solving approach will be used throughout the course. Under the guidance of experienced lab instruc-tors, participants will work in groups of three to plan their ownprotocols, analyze data, and interpret results. A student-teacherratio not greater than 8:1 will be maintained and the faculty coor-dinators will be present throughout the course.Special RecognitionWe would like to recognize and thank Dr. Dennis Willows,Director of University of Washington?s Friday Harbor Labora-tories, for allowing us to use FHL facilities for collection andprocessing of the Aequorea jellyfish used in this course. ?I am sending you another note of thanksto tell you again how much I fondly rememberthe course at Rutgers. I recently attended acourse given by another company which greatlyilluminated the great job that you are doing withyour courses, in terms of providing expert infor-mation in an environment that because of itsenjoyable atmosphere is highly conductive tolearning. Keep up the good work?Bruce Baldwin, Eli Lilly & CompanyCourse LocationThe course will be held on the School of Environmental and BiologicalSciences Campus, New Brunswick, NJ. A map with written direc-tions to the course site will be included with your confirmation ofregistration. If you do not receive your confirmation ten days beforethe course starts, please call our Registration Desk at (732) 932-9763ext. 225 or 216 By Phone9:00 a.m. ? 4:00 p.m., Monday ? FridayRegistration Desk: (732) 932-9763 ext. 225 or 216""""" By MailSend check or money order (payable to Rutgers, the State University) to:Registration DeskRutgers UniversitySchool of Environmental and Biological Sciences (CREBB)Department of Biochemistry and Microbiology76 Lipman DriveNew Brunswick, NJ 08901-8525By Credit CardVisa or MasterCard onlyCall Registration Desk (732) 932-9763 ext. 225 or 216 By Fax(732) 932-3633, 24 hoursPlease fax a copy of check or money order with fax registration." Via the Internethttp://www.rci.rutgers.edu/~crebb/registerprotein.htmlWays to RegisterFor information on travel and lodging, please contact: GloriaKierniesky at (732) 932-9763 ext. 225 or e-mail her atkierniesky@aesop.rutgers.edu, or phone the Rutgers InformationService at (732) 932-INFO. The following Rutgers website containsinformation on discounted travel and lodging:http://www.rutravel.rutgers.edu/discountprograms.htmlGGGGRefundsYou may withdraw from this course with a full refund (less a $100processing fee) provided our office is notified at least five (5) fullbusiness days prior to the start of the course. Beyond that time, youmay be responsible for the full tuition fee if you register but do notattend. Substitutions are encouraged.CREBB reserves the right to cancel this course due to insufficientenrollment.Registration InformationThe tuition fee for this five and one-half day course is $2,695; registerthree weeks prior to the start of the course and pay only $2,195. Adiscounted fee of $1,995 per person is available if two or moreregistrants from the same company register. There is also a discountedfee of $1,395 for attendees from academia. Fees MUST BE PAIDthree (3) weeks prior to the start of the course to be eligible for thediscounted rates. The tuition fee includes continental breakfasts,lunches, coffee breaks and three dinners, along with all coursematerials and hand-outs.Course FormatCourse participants will extract an easily visualized chromo-protein, the green-fluorescent protein, (Science vol. 263pp. 802-805,1994) from a frozen tissue sample or bacterial cellpellets, clarify the extract, and then concentrate and purify theprotein by ?salting out.? Gel filtration, ion exchange, hydro-phobic interaction, and size exclusion HPLC chromatographywill then be employed to extensively purify the desired protein(GFP) from the crude extract. The unique nature of thisbrilliantly fluorescent protein allows you to follow all phases ofthe purification with a simple hand-held mineral light, enhanc-ing your understanding of each process.The purified protein will be characterized by SDS and nativegel electrophoresis, isoelectric focusing, ion exchangeFPLC, size exclusion HPLC, and Western blotting. Each groupwill prepare a detailed purification table and graphs (homeworkassignments), and will characterize the protein with respect topurity, charge, molecular weight, isoelectric point, uniquespectral features, subunit composition, isoprotein composition,and the chemical nature of the chromogenic peptide.This course integrates lecture and laboratory sessions toprovide a comprehensive learning experience. The coursebegins with an introductory lecture on Sunday afternoon.Everyone is strongly encouraged to attend this session, butparticipants who cannot arrive for the Sunday lecture maybegin the course on Monday morning (at the laboratorylocation).The course concludes Friday afternoon with an interactiveproblem-solving workshop and a molecular modelingworkshop at the Rutgers Structural Biology ComputationalLaboratory (http://cesario.rutgers.edu) facilities in Lipman Hall,Rutgers Out-of-state registrants should try to schedule flighttimes out of Newark International Airport no earlier than 6:00p.m. so they do not miss this session. ?I got far more out of the course than I had even anticipated, and it wascertainly money VERY WELL spent. In that one week, the training was farmore intense and the knowledge I gained more directly applicable to what Iam working on than I could have attained otherwise. I still refer to themanual that you provided. I also appreciated the assistance and knowledgeof your collaborators.?Robert A. Bollinger, Dept. of Radiology, UT Southwestern MedicalCenter ?Dr. Ward was very available and open to questions. He sincerelytried to answer every question asked and was very patient. The TAs wereawesome, very well prepared and eager to help.?Bridget Ferdinand, Manufacturing Associate, Antigenics, L.L. C.Sunday 3:00 p.m. - 8:00 p.m.Registration and Reception 3:00 p.m. - 4:00 p.m.Course Introduction and Overview 4:00 p.m. - 7:00 p.m.Dinner 7:00 p.m. - 8:00 p.m.Monday 7:30 a.m. - 8:00 p.m.¾ Lecture: Protein Structure¾ Laboratory Introduction and Overview¾ Morning Laboratory Exercises: Filtration, Precipitation with Ammonium Sulfate, Centrifugation, Fluorimetric and Spectrophotometric Assays,and Gel Filtration Chromatography¾ Lecture: General Preparative Methods of Protein Purification¾ Evening Laboratory Exercises: Do Biochemical Assays, Plot Gel Filtration Data, Select Ion-Exchange Conditions, Start Dialysis,and Begin Purification TableTuesday 7:30 a.m. - 5:30 p.m.¾ Lecture: Open Column Purification¾ Demonstration: Analytical Gel Filtration¾ Morning Laboratory Exercises: Centrifuge Dialyzed Sample, Begin Ion-Exchange Chromatography¾ Afternoon Laboratory Exercises: Choose HIC Matrix and Eluting Solvent, Assay Ion-Exchange Fractions, Plot DataWednesday 7:30 a.m. - 8:30 p.m.¾ Morning Laboratory Exercises: Run HIC on Peak Ion-Exchange Fractions¾ Lecture: HPLC Theory¾ Morning Laboratory Exercises: Assay HIC Fractions¾ Afternoon Laboratory Exercises: Concentrate and Desalt Samples, Purify by SEC-HPLC and Calculate Molecular Weight, Analyze Pure GFPSpectrally¾ Lecture: Electrophoresis¾ Evening Laboratory Exercises:Demonstration: Tangential Flow Ultrafiltration (Pall Minimate TFF System)Demonstration: Three Phase Purification of Proteins using t-Butanol¾ Interactive Workshops1. Affinity Chromatography of His-tagged Protein on Qiagen Nickel Column2. Western Blotting on the Novex System3. Titration Curve on GE Healthcare “Phast” SystemThursday 7:30 a.m. - 5:30 p.m.¾ Demonstration: Preparative PAGE on BioRad Prep Cell¾ Morning Laboratory Exercises: Pour SDS Running Gel, Load Isoelectric Focusing Gel¾ Demonstration: “Phast” System, Native Gel with Western Blot¾ Lecture: Analysis of Protein Structure¾ Laboratory Exercises (continued): Pour SDS Stacking Gel, Stain IEF Gel¾ Afternoon Laboratory Exercises: Run SDS Gel, Stain and Destain SDS GelsFriday 7:30 a.m. - 4:00 p.m.¾ Lecture: Molecular Biology of GFP or HPLC of Proteins and Peptides¾ Morning Laboratory Exercises: Analyze SDS Gels, Plot SDS Molecular Weight Data, Analyze IEF Gel, Determine Isoelectric Point, Analyze“Phast” IEF, Complete Purification Table¾ Interactive Workshop: Determination of Chromopeptide Structure: Analyze Chromopeptide Purification Data. Discuss and Analyze: (1)Amino Acid Composition Data, (2) Edman Sequencing Data, (3) Pronase/Carboxypeptidase Data (4) DNA Sequence Data and (5) ModelCompound Analysis.¾ Molecular modeling workshop: Rutgers Structural Biology Computational LaboratoryNote: Breakfast and Lunch included Monday - Friday. Dinners provided Sunday, Monday and Wednesday.Course OutlineUpcoming Biotech Course offered by CREBBBiochemical SeparationsIntroduction to Laboratory Techniques(3 days, lecture/demonstration)This course provides an introduction to techniques used to isolate,purify and characterize biological macromolecules. Emphasis is placedon techniques used in protein isolation and purification ? both fromnative and recombinant sources. Techniques include extraction, filtra-tion, centrifugation, column chromatography including HPLC, elec-trophoresis, isoelectric focusing, western blotting and capillary zoneelectrophoresis. This is an introductory level course designed for sci-entists having limited experience with laboratory separations.Course Cost: $995September 19 - 21, 2008 (Fri - Sun)2009 Dates TBAWebsite:http://www.rci.rutgers.edu/~crebb/biosep.html New for 2008 ##########Tutorials in Protein Purification(3 days, hands-on instruction in Downstream Processing)In an intimate, small group laboratory setting, learn the details ofprotein purification from an expert. Join Professor William Ward ashe leads you through the step-by-step purification of a model pro-tein, hands-on from start to finish. Under his constant supervision,you will prepare a crude extract, administer several batch purifica-t ion steps, and then achieve purity in a series of high resolutionchromatography steps. You will then judge purity of the final prod-uct in comparison with your crude extract by calculating specificactivity and by analyzing your product by size exclusion HPLC and/or SDS gel electrophoresis.Course Cost: $995June 27 - 29, 2008 (Fri - Sun)October 10 -12, 2008 (Fri - Sun)2009 Dates TBAWebsite:http://www.rci.rutgers.edu/~crebb/Tutorials.htmlEarly Registration: If you register three weeks prior to the start of thecourse you pay only $895.Discounted Fee: A discounted fee of $850 per person is available if twoor more registrants from the same company register.The tuition fee includes continental breakfasts, lunches, and coffee breaksalong with all course materials and hand-outs.Techniques and Instruments You Will Usev Tissue Homogenizers (Omni)v Filtration Devices including Tangential FlowUltrafiltration (Millipore, Sartorius, Vivascience, Pall)v Refrigerated (Sorvall) Centrifugesv Recording UV-Vis Spectrophotometers (Spectronic, Cary)v Filter Fluorometers (Turner Designs, Hoefer)v Chromatography: Gel Filtration, Ion-Exchange, & Hydro-phobic Interaction (GE Healthcare, BioRad)v Immobilized Metal Ion Affinity Chromatography(Qiagen, BD Biosciences/Clontech)v Fraction Collectors (Gilson, Isco)v Column Monitorsv Electrophoresis (SDS, native and isoelectricfocusing) (BioRad, Invitrogen/Novex, GE Healthcare)v Western Blotting (Invitrogen/Novex)v Coomassie Blue and Silver Staining (Sigma)v Ion Exchange HPLC (Applied Biosystems)v Size Exclusion HPLC (Thermo Electron Corporation,Phenomenex)v GE Healthcare Phast Electrophoresis Systemv Molecular Modelingv Amino Acid Analysis*v Automated Edman Sequencing*v DNA Sequence Analysis*v Peptide Mapping*v Mass Spectral Analysis**Introduced in problem-solving session on Friday ?I was extremely impressed with thequality of the course and caliber of allinstructors ... best course like this I?ve beento ... Thank you for a wonderful learningopportunity and environment ...?Margo Zanotto, Manufacturing Associ-ate, Biotech, Baxter BiotechCorporate Support: Equipment, materialsand laboratory supplies from the followingcompanies are used in this course. Wesincerely appreciate their support.NameFirst Name For NametagEmployerJob TitleMailing AddressWork PhoneHome PhoneFax No.E-mail AddressRutgers University, School of Environmental andBiological Sciences (CREBB)Department of Biochemistry and Microbiology76 Lipman DriveNew Brunswick, NJ 08901-8525GFP GuideD&MD Publications is happy to extend a10% discount to course registrants for ?AGuide to Green-Fluorescent Protein:Applications in Cell Biology and DrugDiscovery.? To order the Guide, please contact D&MD?sCustomer Service Department atcust.serv@drugandmarket.com or call+1 (508) 616-5566 and mention sourcecode ?9133-10%PP?.Tuition Fee:$2,195 (Early Registration)$1,995 (Per Person for MultipleEarly Registration of 2 or more attendeesfrom the same company)$1,395 (Academic Early Registration)$2,695 (Regular Registration)NOTE: Fees MUST BE PAID3 weeks prior to the start of thecourse for all discounted rates.#Check or Money Orderenclosed in the amount of$______________#Visa or MasterCard# _________________________Exp.Date:____________Protein Purification$June 1 - June 6, 2008$July 13 - July 18, 2008$July 27 - August 1, 2008$January 11 - January 16, 2009$March 15 - March 20, 2009ProteinPurification:Principles and PracticeNonprofit OrganizationU.S. Postage PaidNew Brunswick, NJPermit # 1296Rutgers University, School of Environmental andBiological Sciences (CREBB)Department of Biochemistry and Microbiology76 Lipman DriveNew Brunswick, NJ 08901-8525