lab for engineering in infection & immunitylab for engineering in infection & immunity...
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Lab for engineering in infection & immunity Engineering proteins to control immune responses and bacterial pathogenesis
Jennifer Maynard, Dept. Chemical Engineering
Bioreactor Cell as reactor Products
Interactions
Catalytic
Self-assemblyProteins
Mechanicalwork
Therapeutics
Complexmachines
Signaling
GenesOn/off
Structural
Power of engineering
Understand underlying biology such that we can develop design rules to predictably manipulate systems
Basic science Applied science
I. Designer TCR technologyDiagnostics - track pMHCs in response toDisease & vaccination
Immunotherapeutics - target cancer/ virallyInfected cells, alter autoimmune disease
Basic science- ability to study properties in Vitro; low affinity techniques; design rules
GoalsControl affinity and specificity
ChallengesLow affinity (1-50 uM) - hard to study interactionRules for recognition unclear
Humancell
peptide MHC
T cell
TCR
0
50
100
150
200
250
300
mAU
0.0 5.0 10.0 15.0 ml
S75 SizeExclusion
0
20
40
60
80
100
120
140
160mAU
5.0 10.0 15.0 20.0 25.0 ml
Anion Exchange
DO
11.1
0 sc
TC
R
MHC
TCR 1 2 3 4 5 6 7 8 9 10Wild-type C A A S P N Y N V L Y
1721 C A D D Q S N R Y17210 C A A G H R C N Y1322 C A A * Y T R S P L Y1328 C A A S S F T P R L Y1744 C A A A L N Y H S L Y1746 C A A P R T A F N L Y1761 C A A S T N WH N L Y2343 C A A V P N Y H L L Y2144 C A A S Y A Y H Q L Y2146 C A A T P W L S R L Y
176-1, 0.2 nM 235-1, 20 nM DO11.10, 40 nM
Production
Control affinity
1
3
2
OVA 323-339/ I-Ad pMHC
DO11.10scTCR
1
3
2
Ac1-11 MBP/ I-Au pMHC
172.10scTCR
OVA 323-339/ I-Au pMHC
172.10scTCR
with DO11.10CDR 3α and β
1
3
2
?
Control specificity
Vβ8.2
Vα13.1
DO11.10 model T cell/ cancerDemonstrate the utility of customized
TCRs through in vitrocompetition experiments, in vivotumor targeting
172.10 is an autoimmune TCRTarget: Myelin Basic ProteinAc1-11
Isolate high affinity mutant fortargeted immunosuppression inmouse model of multiplesclerosis
I. Immediate applications
Structural Determination: NIH Structural InitiativeProvide insight to protein mechanism for vaccine and drug design
Bioseparations: Control of Down Stream Process (ex Insulin)Controlled Drug Delivery
II. Building protein crystalsGoals:Understanding Latticecontacts:
weak interactions,H-bonds,2nd virial Coeff.
Hypercrystallizable Ab forScaffold
Challenges:Crystallization: Nucleationand GrowthLow % of proteinscrystallized
II. Building protein crystalsCrystal lattice formation Crystal contacts: repeating units
III. Reverse engineering of pathogens
Bacteria Cellinvasin
FACs labeling of functional,surface Expressed Invasin
Model for vaccine Delivery system
III. Whooping coughToxin-mediated disease-- immunostimulatory and inhibitory aspects of PTx?-- neutralizing epitopes?
1B7
PTx-1B7 interaction
Bordetella pertussis Toxin mechanism
Power of engineeringUnderstand underlying biology such that we can develop design rules to predictably manipulate systems
Basic science Applied science
Focus on TCRs and Antibody fragments with the following techniquesRecombinant TechniquesProtein Expression in Eukaryotes and Bacteria (E. Coli)Protein CharacterizationProtein CrystallizationCell Culture
Maynard laboratory will be offering 2 graduate research assistant Positions in Fall 2009 in the areas of T cell receptors and protein expression
Contact: maynard@che.utexas.edu
Members of the lab from left to right: Ben Roy, Ryan Myhre, Tarik Khan, Jamie Sutherland and Jen Pai
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