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