1st Annual Unither Conference, April 2, 2008

Self-Assembly of NanoBiomaterials for Tissue Engineering

Qian Wang
Department of Chemistry and Biochemistry Nanocenter University of South Carolina

Nanomedicine, 2006, 137 Chem. Commun. 2007, 1453 Optics Commun. 2005, 366 TL, 2005, 1691; JACS, 2007, ASAP Bioconj. Chem, 2007, 852 Bioorg. Chem. 2008, in print See also: ACIE, 2002, 459; Chem. & Biol. 2002, 805 & 813; Biomacromolecules, 2003, 472; Bioconjugate Chem. 2003, 38; Biomacromolecules, 38 Chembiochem, 2003, 1348.

Chem. Commun. 2006, 3019; ACIE, 2005, 2420; Science, 2006, 312, 1631; Nature 2005, 434, 55; Micro & Nano Lett., 2006, 1; Langmuir, 2007, 6719; NanoBiotechnoloty, 2007, 31; Langmuir, 2007, 11157; Nano Lett, 2007, 7, 3729

Chembiochem, 2008, 9, 519 Acta Biomater. 2008, in print

Structural Feature of Icosahedral Cowpea Mosaic Virus (CPMV)

Non-envelop, icosahedra Diameter = 31.4 nm Two protein per asymmetric units 60 copies of asymmetric units MW (capsid) = 3.6 x 106 D Two RNAs as genome MW (with RNA) = 5.9 x 106 D

CPMV

TYMV

T. Lin, Z. Chen, R. Usha, C.V. Stauffacher, J. Dai, T. Schmidt, J.E.

Johnson: Virology 1999, 265, 20

Structural Feature of Helical Tobacco Mosaic Virus (TMV)

mrsec.wisc.edu

Availability Stability and safety Well-defined structures Identical in size and shape Genetically programmable Robust chemistry

Plant Virus Isolation

0.59 0.49

TMV at 260nm TMV at 280nm

absorbance (a.u.)

0.39 0.29 0.19 0.09

Homogenize infected leaves

-0.01 0 5 10 15 20 25 30

20% sucrose cushion Differential centrifugation

volume (mL)

Extraction

PEG8K and NaCl Triton X-100

Plant homogenate 2 sq ft

Aqueous layer 0.5-1 kg tobacco leaves

Organic solvent 1-2 g particle (>97% pure) < $200 per gram

Representative Virus Based Nanomaterials Development

CPMV / NV : Finn & Johnson

CCMV: Douglas, Young, Zlotnick

TMV: Mann, Douglas, Culver, Francis, Wang, etc

CPMV: Ratna, Soto, Blum & Coworkers

MS2/TMV: Francis

RCNMV: Franzen

TYMV: Wang

Bionanoparticles Templated Functional Materials

Catalytic Mesoporous Scaffold Functional Membrane Conductive Nanowire

Bionanoparticles

targeting units

In-capsid probes antibody peptide insertions

Biocomposite Materials

Drug Delivery Vehicle Artificial Cell

Qian Wang

Cell-Culturing on Self-Assembled Plant Viruses

Chem. Commun. (2006) 3019 NanoBiotechnology (2007) in print Angew. Chem. Int. Ed. (2005) 2420 Science (2006) 312, 1631 Nature (2005) 434, 55 Micro & Nano Lett. (2006) 1, 1

Controlled Assembly: 1D nanofiber 2D assay 3D matrix

Virus Chemistry: Production and characterization Selective functionalization

Cell Culture: Fibroblast cells Bone Marrow Stems cells Cardiac Myocytes

Four Basic Ways of BNP Functionalization

Conventional bioconjugation chemistry Genetic Modification Post-functionalization: Click Reaction In situ polymerization Surface coating via non-covalent interactions

Conventional Bioconjugation Chemistry for BNP Functionalization

R H N O N H O R N H O R O z(HOOC) NH R-NH2 R-OH R-NH-NH2 O O R2 H N R1 N N R2

Glu/Asp Virus

Arg

R1

NH NH2 x

NCS

H N S

H N R

O (HS)m O S R N O O R N H S R N H O Br (I) R NH N H R S S R S S N HO N R S S O R S S CH3 O R N O NaO3S R N3 O N O O UV :N R N O O (NH2)n

Lys

AE

H N R O

Cys
HO y

X R R2 N (F) Cl N R1 O N

H N Y R R2 H N N N N R1 N CH R Reduction H N CH2 R

Tyr

NH R N R N R

O S O

AE =
S R O S O

N O O

H N

H 3C

CH3 N CH3

R = Functional groups X = -COCl; -SO2Cl; -CON3

Y = -CO-, -SO2-

Finn and Johnson, Angew. Chem. Int. Ed. 2002, 41, 459; Andrew Lee: Nanomedicine, 2006, 137

Selective reactions on CPMV Cys Chimeras

O O N O O Biotin derivative HO NH H N O H HN O S O H NH (H2N)60 S O N Biotin 60

OH HO AcHN O N O

OH O O N H H N O OH

O (H2N)60 S O N Tn-Antigen 60

+
(H2N)60 (SH)60

Tn-Antigen Cys-CPMV

Hannah Barnhill, collaboration with Dr. Xuefei Huang and M. G. Finn

VBCCys
O

VEFCys

R3P PR3 N Au PR3 PR3 R3P PR3 O R3P NanogoldR

Finn & Johnson; ACIE, 2002

Finn & Johnson; ACIE, 2002

Virus: The Programmable Supramolecular Platform

(CH2CH2O)n CH3

DNAs

(CH2CH2O)n

Drugs

Polymer
HOOC
N
N

O C N H
COOH

Proteins

HOOC

COOH

MRI Ligands
O
O
NH

HN

NH

OH

HOOH

S
O

Fluorescent Dyes

O O
O NHAc

O
O

biotin

OH

Carbohydrates

JACS 2007, 7799; Nanomedicine, 2006, 137; Chem. Commun. 2007, 1453; Optics Commun. 2005, 366; Tetrahedron Lett. 2005, 1691; Bioconj. Chem. 2007, 853; Nanobiotechnology, 2007, accepted. See also: Angew. Chem. Int. Ed. 2002, 41, 459; Chem. & Biol. 2002, 9, 805 & 813; Biomacromolecules, 2003, 4, 472; Bioconjugate Chem. 2003, 14, 38; Nano Lett. 2003, 3, 883; Chembiochem, 2003, 4, 1348. 38

Click on TMV - A Modular Process

Mike Bruckman

2D Assembly at Liquid-Liquid Interface

Organic solvents

Oil droplets stabilized by bioparticles

interface

BNPs

(A)
shaking crosslinking

organic polymer

BNPs BNP assembly

(B)

Water Oil

Water Oil

crosslinking

In collaboration with Russell and Thiyagarajan

2D Assembly at Liquid-Liquid Interface

Su Long, Gagandeep (in collaboration with Prof. T. P. Russell, UMass)

40m

Interfacial Assembly Using Polymer as Organic Phase

Viruses DMF/Water Dialysis

P4VP

Tao Li

Hypothesized Pathway of Hierarchical 1D Assembly of TMV

1D assembly

surface recognition

Po l ym eriz atio n

2D assembly

AFM & TEM of Tobacco Mosaic Virus

Zhongwei Niu

Synthesis of TMV/Polyaniline Composite

TEM Images of Polyaniline/TMV Nanofibers

Zhongwei

AFM Images of One Dimensional Fibers

Zhongwei

Conductive Nanowires

Zhongwei Niu (in collaboration with G. Koley and C. Li)

Wang Group: Dr. Zhongwei Niu Dr. Cline le Droumaguet Dr. Jianhua ROng Michael Bruckman Siqi Li L. Andrew Lee Gagandeep Kaur Tao Li Kai Li Jerry Oxsher Laying, Eve, Sharmi, Lily Fang Xie (Ph.D.) Hannah Barnhill (Ph.D.) Brandon Cash (M.S.) Su Long (M.S.) Swat Kabisatpathy Dr. Seok-Joon Lee Dr. K. Sivakumar Dr. Srinivas Chitaboina Dr. Venkata S. Kotakadi Dr. Qingbing Zeng Dr. Kris Varazo Daiki Kyotani Lisa Alexander Preston Thompson Other Undergraduate Students

Acknowledgments
BNP Assembly: Prof. Tom Russell Yao Lin Dr. Alex Bker Jinbo He Dr. Andreas Fery Prof. Anna Balazs Prof. T. Dinsmore Dr. P. Thiyagarajan Dr. Byeongdu Lee Dr. Lin Yang Prof. Cathy Murphy Prof. Ken Shimizu Prof. J. Lavigne Prof. H. zur Loye Cell Study Prof. Yu-Li Wang Prof. J. Potts Prof. Z. Su Prof. S. Kistler Prof. E. Jabbari Protein Binding Study Prof. David Tirrell Prof. Lukasz Lebioda Virus Propagation: Prof. Theo Dreher Prof. Rick Nelson Prof. Vicki Vance Click Related Reaction: Prof. K. Barry Sharpless Prof. M.G. Finn Prof. Valery Fokin Prof. Greg Tew Prof. Todd Emrick Prof. Dan Reger Cyanine Dye Synthesis: Prof. He Tian Wenhai Zhan $$$ Alfred Sloan Foundation NSF-CAREER NSF-NER/NSF-DMR DOD-MURI / DOD-DURIP ARO-BCRP W. M. Keck Foundation USC Research Foundation USC NanoCenter SC EPSCoR Office & SCCC