Tailoring carbon nanotube surfaces with glyconanorings: New bionanomaterials with specific lectin affinity

Tailo ing ca bon nano ube su aces wi h glyconano ings: new bionanoma e ials wi h specific lec in affini yw Nou eddine Khia ,* a Manuel Pe nia Leal, a Rachid Baa i,* b Ch is ine Ruhlmann, c Cha les Mioskowski,z b Pa ick Schul z c and Inmaculada Fe na ´ndez d Recei ed (in Camb idge, UK) 9 h Ma ch 2009, Accep ed 1s May 2009 Fi s published as an Ad ance A icle on he web 2nd June 2009 DOI: 10.1039/b904717d Rema kably s able, wa e -soluble glyconano ing-coa ed SWCNTs we e p epa ed by sel o ganiza ion and pho o- polyme iza ion o neu al diace ylene-based glycolipids on he nano ube su ace; he nanocons uc s a e able o engage in specific ligand–lec in in e ac ions in a simila way o glycoconjuga es on cell memb anes. Endowed wi h unique size, shape and physical p ope ies, single wall ca bon nano ubes (SWCNTs) a e cu en ly being ac i ely in es iga ed as a ehicle o he in i o sma deli e y o biologically ele an molecules, as nanome ic senso s, and po en ially, o cance ea men . 1 In e es ingly, ecen in es iga ions, including an in i o pilo s udy, ha e concluded ha con enien ly unc ionalized wa e -soluble SWCNTs a e non- oxic. 2 The e o e, he de elopmen o efficien s a egies o he cons uc ion o wa e -soluble, biocompa ible CNTs is a challenge o pa amoun impo ance. 3,4 Owing o hei high su ace a ea, bioenginee ed CNTs can be used o expose a la ge numbe o biomolecules, in a manne ha is simila o he species in ol ed in hose c i ical biological p ocesses ha a ou mul i alen ligand– ecep o in e ac ions. 5 A pa adigma ic example o such e en s a e hose media ed by ca bohyd a e–p o ein in e ac ions, 4 which include cell adhesion, inflamma ion, umou cell me as asis, and pa hogenic in ec ions. 6 Building on ou in e es in he syn hesis o CNT-based bionanoma e ials, 7 he ein, we epo a new bo om-up app oach o he p epa a ion o ema kably s able wa e -soluble SWCNT–ca bohyd a e nanohyb ids wi h a nanoabacus opology and wi h specific lec in affini y. Ou a ionally designed app oach o he desi ed glyconano ing- coa ed SWCNTs, Fig. 1, is based on he p e iously epo ed sel o ganiza ion a ound ca bon nano ube su aces o cha ged anionic su ac an s, 7a,8 and on he known selec i e and smoo h pho opolyme iza ion o diace ylene-based glycolipids. 9 Compa ed o o he amphiphiles, glycolipids a e cha ac e ized by a complexi y o in e ac ions among he suga g oups, which add fine de ails o he hyd ophilic–hyd ophobic balance go e ning hei sel assembly. 10 Thus, o ailo he p ope ies o he suga -based biosu ac an o he desi ed sel o ganiza ion on he nano ube, a diace ylene-based neoglycolipid Iwas designed, which exhibi s a 25 ca bon-based hyd ophobic ail o an efficien an de Waals in e ac ion wi h he CNT, wi h a a iable space and suga g oup o fine uning o he hyd ophilic–hyd ophobic balance o he molecule. Neu al de e gen s 7–11 used in his s udy (Scheme 1), we e ob ained by a new, con e gen , and modula s a egy, s a ing om (2-aminoe hyl)-pe -O-ace yla ed-1- hio-glycosides 1,2, and 3, o which a one s ep app oach has been de eloped (see ESIw). Sonica ion o SWCNTs in he p esence o a homogeneous aqueous solu ion o Lac-7affo ded uns able agg ega es as e idenced by apid sedimen a ion o he ca bon nano ubes in he medium. This esul e eals ha lipid Lac-7is unable o sel o ganize on he nano ube side-wall, mos p obably due o i s in insically un a o able hyd ophilic–hyd ophobic balance. Fig. 1 O e iew o he glyconano ing s a egy. Scheme 1 S uc u es o (2-aminoe hyl)-pe -O-ace yla ed-1- hio-lac oside 1, cellobioside 2, and mannoside 3, he bi unc ional space s 5–6, and he diace ylenic acid 4used o he syn hesis o diace ylene-based Lac-7, Lac-8, Cellob-9, Man-10, and Man-11. a Ins i u o de In es igaciones Quı´micas, C.S.I.C-Uni e sidad de Se illa, c/. Ame ´ ico Vespucio, 49, Isla de la Ca uja, 41092 Se illa, Spain. E-mail: [email p o ec ed]; Fax: +34 95 44600565; Tel: +34 95 448 9559 b Uni e si e ´de S asbou g, Facul e ´de Pha macie CNRS UMR 7199 Labo a oi e de Chimie des Sys e `mes Fonc ionnels, 74, ou e du Rhin, 67401 Illki ch-G affens aden, F ance E-mail: [email p o ec ed] c Ins i u de Ge ´ne ´ ique e de Biologie Mole ´culai e e Cellulai e, CNRS/INSERM/ULP, 1 ue Lau en F ies, BP163, F-67404 Illki ch Cedex, F ance d Depa amen o de Quı´mica O ga ´nica y Fa mace ´u ica, Facul ad de Fa macia, Uni e sidad de Se illa, 41012 Se illa, Spain wElec onic Supplemen a y In o ma ion (ESI) a ailable: Expe imen al p ocedu es, lec in binding assays, NMR, IR and Raman spec oscopies and addi ional TEM images. See DOI: 10.1039/b904717d zDeceased. Dedica ed o he memo y o D Cha les Mioskowski. This jou nal is cThe Royal Socie y o Chemis y 2009 Chem. Commun., 2009, 4121–4123 |4121 COMMUNICATION www. sc.o g/chemcomm |ChemComm Published on 02 June 2009. Downloaded by G al Uni e sidad Se illa on 22/06/2016 16:23:44. View A icle Online / Jou nal Homepage / Table o Con en s o his issue In o de o alida e his hypo hesis, he mo e hyd ophilic lac ose based su ac an Lac-8was syn hesized. This has been achie ed by he inco po a ion o he bi unc ional e hylene glycol de i ed space 5, ob ained om e ae hylene glycol in h ee s eps (see ESIw) ia amide bond linkage, o he (2-aminoe hyl)-2,20,3,30,40,6,60-hep a-O-ace yl-1- hio-b-lac oside 1. A e S audinge azide educ ion, a syn he ic ou e simila o ha used o Lac-7, affo ded glycolipid Lac-8in good yield. Consequen ly, by simply mixing he neu al lipid Lac-8wi h CNT in wa e wi hou any addi i e, ollowed by sonica ion o 30 min, a black solu ion which emained s able o mon hs was ob ained (Fig. 2A, ial 2). A simila s able black suspension was ob ained when glycolipid Cellob-9de i ed om cellobiose disaccha ide was used wi h he same SWCNTs (Fig. 2A, ial 3). These esul s show ha an inc ease in he hyd ophilic–hyd ophobic balance o he glycolipids, by adding a poly(e hylene glycol) (PEG) chain, is c ucial o he unc ionaliza ion o SWCNTs and he o ma ion o s able nanocons uc s in pu e aqueous condi ions. This obse a ion is co obo a ed by he ac ha : (i) a dec ease o he hyd ophilic– hyd ophobic balance o he su ac an by using a monosaccha ide such as in Man-10, ga e exclusi ely uns able agg ega es ha p ecipi a ed quickly in wa e , and (ii) he use o neoglycolipid Man-11 wi h he same suga epi ope bu a longe hyd ophilic space 6allows he ema kable solubiliza ion and unc ionaliza ion o ca bon nano ubes in pu e wa e . The mac oscopic obse a ion o a s able black suspension is a esul o he change in he na i e hyd ophobici y o he nano ube side wall, becoming mo e hyd ophilic, due o he unc ionaliza ion o he SWCNT’s su ace wi h glycolipids and he exposu e o he suga epi opes o he pola wa e phase. The cha ac e iza ion o SWCNT–neoglycolipid nanocons uc s was ca ied ou by ansmission elec on mic oscopy (TEM), and nuclea magne ic esonance (NMR), in a- ed (IR) and Raman spec oscopies (see ESIw). TEM images o na i e SWCNTs and SWCNTs-coa ed wi h neoglycolipid Lac-8a e gi en in Fig. 2B and 2C, espec i ely. As a consequence o he hyd ophobic in e ac ions, an de Waals o ces, and p-s acking among indi idual nano ubes, as p oduced SWCNTs usually exis as dense bundles o opes ha a e deeply in e connec ed, Fig. 2B. 11 Qui e ema kably, a simple unc ionaliza ion wi h neoglycolipids Lac-8, Cellob-9 o Man-11 in pu e wa e and s ic ly neu al condi ions, allowed he CNTs packages o ex olia e p oducing small mainly indi idual suga -coa ed nano ubes, as e idenced by TEM analysis (Fig. 2C, D and E). In e es ingly, he TEM images show ha he su ace o he nano ube is comple ely co e ed by s ia ions o hemi-micelles, ha can slide on he nano ube in a simila way o beads on a single wi e in an an ique abacus. Simila associa ions we e p e iously epo ed o cha ged and zwi e ionic amphiphilic lipids, 7,8 bu as a as we know his is he fi s ime whe e such o ganiza ion is epo ed o a neu al, non-ionic su ac an in pu e wa e . 12 While up o h ee mechanisms we e p oposed o explain he dispe sion o CNTs wi h su ac an s, 13 he obse ed pe iodic s ia ions sugges ha he mic oscopic binding mode is he hal cylinde mode. 7a In e es ingly, unde such sup amolecula o ganiza ion i is well documen ed ha diace ylene lipids unde go efficien and clean polyme iza ion ia a 1,4-addi ion eac ion o o m al e na ing ene-yne polyme chains upon ligh i adia ion a 254 nm, 9 affo ding s ong polyme ized glyconano- ings a ound he su ace nano ubes as shown in Fig. 3. 14 A e sonica ion o SWCNTs in he p esence o aqueous solu ion o Lac-8as be o e, he mix u e was hen i adia ed by a labo a o y UV lamp (254 nm) o 12 h in o de o p omo e he polyme iza ion o he diyne unc ional g oups a ound he nano ube. The high s abili y o he polyme ized glyconano ings was asce ained by compa ison wi h ha o he non-polyme ized coun e pa nanohyb ids. While he la e p ecipi a ed a e h ee mon hs he o me ones emained s able o a leas six mon hs. Addi ionally, he s able suspension o CNTs unc ionalized wi h polyme ized glyconano ings emained unchanged a e washing wi h me hanol, hea ing in wa e a 80 1C o one week, and when dilu ed in Hepes buffe (20 mM a pH 7.5)—in con as wi h he non-polyme ized CNT–glycolipid nanocons uc s and o mos o he su ac an -based CNT-solubiliza ion sys ems de eloped so a (see ESIw). These esul s indica e ha he SWCNTs a e coa ed wi h polyme ized polydiace ylene-based glyconano ings (PDA-GNR), o ming s able, wa e -soluble mul icomposi es namely SWCNT-PDA-GNR-Lac-8. I is wo h men ioning ha his ea u e ex ends he u ili y o ou bionanoma e ials, since polydiace ylene- e he ed biomolecules ha e ecen ly ecei ed a g ea deal o a en ion as biosenso s, due o hei unique p ope ies upon ex e nal s imula ion. 9b A cen al issue o his s udy was o demons a e he abili y o such suga epi ope-bea ing bionanoma e ials o engage in specific in e ac ions wi h p o ein ecep o s. Theo e ically, each polyme ized nano ing on he nano ube su ace is su ounded by a la ge numbe o suga epi opes, much like he glycocalyx Fig. 2 (A) Pho og aphs o ials con aining aqueous solu ion o : 1. SWCNTs, 2. SWCNT-Lac-8, 3. SWCNT-Cellob-9, 4. SWCNTs- Man-11. TEM images o he nanocons uc s nega i ely cha ged wi h u anyl ace a e: (B) as p oduced SWCNTs, (C) SWCNT–Lac-8, (D) SWCNT–Cellob-9, (E) SWCNT–Man-11. Fig. 3 Fo ma ion o SWCNTs coa ed wi h polydiace ylene-based glyconano ings in pu e wa e . 4122 |Chem. Commun., 2009, 4121–4123 This jou nal is cThe Royal Socie y o Chemis y 2009 Published on 02 June 2009. Downloaded by G al Uni e sidad Se illa on 22/06/2016 16:23:44. View A icle Online a he cell memb ane su ace, and can ac as ligands o specific ecep o s. The e o e he SWCNT-PDA-GNR-Lac-8exhibi ing lac ose esidues on he nano ube’s su ace could be ecognized by a lac ose-specific ecep o such as Peanu agglu inin (PNA) om A achis hypogaea. 15 In o de o confi m his hypo hesis, we incuba ed SWCNT-PDA-GNR-Lac-8, wi h PNA in Hepes buffe a pH 7.4 o 1 hou , and he esul o he in e ac ion was cha ac e ized by TEM. As can be seen om Fig. 4A, he abacus-like geome y was los and he cha ac e is ic s ia ions could no be obse ed. The lec ins co e ed ex ensi ely he unc ionalized nano ubes whose su ace becomes uzzy and om which indi idual lec in molecules can be ound o s ick ou (Fig. 4A, whi e a ows). Whe e hey could be clea ly delinea ed a he edges o he ubes, he sizes o he pa icles a ached o he Lac-8-coa ed SWCNT we e measu ed. A alue o 70 A ˚was ob ained (s=8A ˚,n= 13), which is consis en wi h he size o he 110 kDa PNA homo e ame o which a hyd odynamic adius o 39 A ˚was de e mined by ul acen i uga ion analysis and dynamic ligh sca e ing measu emen s. 16 Addi ionally, he size o he a ached pa icles, Fig. 4A is iden ical o he size o pu ified lec in mac omolecules obse ed by elec on mic oscopy in he absence o nano ubes (see Fig. 4C). As a con ol expe imen o demons a e ha he absence o he s ia ions o med by he glyconano ings is a consequence o he specific in e ac ion be ween he lec in and he nanohyb id, we pe o med he same eac ion be ween he lec in and he glyconanohyb id SWCNT-PDA-GNR-Cellob-9, exposing he cellobiose glycoligand which is no ecognized by PNA. In his case he cha ac e is ic abacus-like geome y can s ill be obse ed (Fig. 4B), as a consequence o he absence o specific in e ac ions be ween PNA lec in and SWCNT-PDA-GNR-Cellob-9. These esul s demons a e ha SWCNTs coa ed wi h glyconano ings can engage in specific molecula ecogni ion wi h a p o ein ecep o and p eclude non-specific p o ein binding. The specific in e ac ions wi h o he biologically ele an species such as cells, bac e ia, and oxins, as well as s udies de o ed o he emo al o hese ascina ing polyme ized glyconano ings (GNR) om he nano ubes su ace, hei eco e y and u he use, a e unde in es iga ion in ou labo a o ies. This wo k was suppo ed by he DGICyT (g an No. CTQ2006-15515-CO2-01 and CTQ2007-61185), he Jun a de Andalucı´a (g an P06-FQM-01852 and P07-FQM-2774), he CNRS (F ance) and CSIC (Egide Picasso 09543XA and PICS P og am 2008). No es and e e ences 1(a) D. Pan a o o, J. B iand, M. P a o and A. Bianco, Chem. Commun., 2004, 16; (b) N. K. W. Kam, T. C. Jessop, P. A. Wende and H. J. Dai, J. Am. Chem. Soc., 2004, 126, 6850; (c)N.K. W. Kam, Z. Liu and H. J. Dai, J. Am. Chem. Soc., 2005, 127, 12492; (d) D. Pan a o o, R. Singh, D. McCa hy, M. E ha d , J.-P. B iand, M. P a o, K. Kos a elos and A. Bianco, Angew. Chem., In . Ed., 2004, 43, 5242; (e) N. K. W. Kam, Z. Liu and H. J. Dai, J. Am. Chem. Soc., 2005, 127, 6021; ( ) S. Lin, G. Keska , Y. Wu, X. Wang, A. S. Moun , S. J. Klaine, J. M. Moo e, A. M. Rao and P. C. Ke, Appl. Phys. Le ., 2006, 89, 143118; (g) N. K. W. Kam, M. O’Connell, J. A. Wisdom and H. J. Dai, P oc. Na l. Acad. Sci. U. S. A., 2005, 102, 11600. 2(a) R. Singh, D. Pan a o o, L. Lace da, G. Pas o in, C. Klump, M. P a o, K. Kos a elos and A. Bianco, P oc. Na l. Acad. Sci. U. S. A., 2006, 103, 3357; (b) M. L. Schippe , N. Nakayama- Ra ch o d, C. Da is, N. K. W. Kam, P. Chu, Z. Liu, X. Sun and H. J. Dai, Na . Nano echnol., 2008, 3, 216. 3(a) D. Tasis, N. Tagma a chis, N. Bianco and M. P a o, Chem. Re ., 2006, 106, 1105; (b) X. Peng and S. S. Wong, Ad . Ma e ., 2009, 21, 625 and e e ences ci ed he ein. 4(a) A. S a , D. W. S eue man, J. R. Hea h and F. S oda , Angew. Chem., In . Ed., 2002, 41, 2508; (b) T. Hasegawa, T. Fujisawa, M. Numa a, M. Umeda, T. Ma sumo o, T. Kimu a, S. Okumu a, K. Saku ai and S. Shinkai, Chem. Commun., 2004, 2150; (c) H. Wang, L. Gu, Y. Lin, F. Lu, M. Meziani, P. G. Luo, W. Wang, L. Cao and Y. P. Sun, J. Am. Chem. Soc., 2006, 128, 13364; (d) P. Wu, X. Chen, N. Hu, U. C. Tam, O. Blix , A. Ze l and C. R. Be ozzi, Angew. Chem., In . Ed., 2008, 47, 5022. 5 M. Mamen, S. K. Choi and G. M. Whi eseides, Angew. Chem., In . Ed., 1998, 37, 2754. 6(a)Y.C.LeeandR.T.Lee,Acc. Chem. Res., 1995, 28, 321; (b)C.R.Be ozziandL.L.Kiessling,Science, 2001, 291, 2357; (c)J.Rojo,V.Dia ´z, J. M. de la Fuen e, I. Segu a, A. G. Ba ien os, H. H. Riese, A. Be na d and S. Penade ´s, ChemBioChem, 2004, 5, 291. 7(a) C. Richa d, F. Bala oine, P. Schul z, T. W. Ebbesen and C. Mioskowski, Science, 2003, 300, 775; (b) C. Thau in, S. Rickling, P. Schul z, H. Celia, S. Meunie and C. Mioskowski, Na . Nano echnol., 2008, 3, 743; (c) M. Assali, M. P. Leal, I. Fe na ´ndez, R. Baa i, C. Mioskowski and N. Khia , So Ma e , 2009, 5, 948. 8(a) M. J. O’Connell, P. Boul, C. E icson, C. Huffman, E. Wang, C. Ha oz, J. Kupe , K. D. Tou , L. M. Ausman and R. E. Smalley, Chem. Phys. Le ., 2001, 342, 265; (b) K. Yu ekli, C. A. Mi chell and R. K ishnamoo i, J. Am. Chem. Soc., 2004, 126, 9902; (c) Y. Wu, J. S. Hudson, Q. Lu, J. M. Moo e, A. S. Moun , A. M. Rao, E. Alexo and P. C. Ke, J. Phys. Chem. B, 2006, 110, 2475. 9(a) D. H. Cha ych, J. O. Nagy, W. Spe ak and M. D. Bedna ski, Science, 1993, 261, 585; (b) D. J. Ahn and J.-M. Kim, Acc. Chem. Res., 2008, 41, 805 and e e ences he ein. 10 M. Co i, L. Can u` , P. B occa and E. Del Fa e o, Cu . Opin. Colloid In e ace Sci., 2007, 12, 148. 11 P. M. Ajayan, Chem. Re ., 1999, 99, 1787. 12 E. J. Wallace and M. S. P. Sansom, Nano Le ., 2007, 7, 1923 and e e ences he ein. 13 (a) K. Yu ekli, C. A. Mi chell and R. K ishnamoo i, J. Am. Chem. Soc., 2004, 126, 9902; (b) M. J. O’Connell, P. Boul, C. E icson, C. Huffman, E. Wang, C. Ha oz, J. Kupe , K. D. Tou , L. M. Ausman and R. E. Smalley, Chem. Phys. Le ., 2001, 342, 265; (c) L. Vaisman, H. D. Wagne and G. Ma om, Ad . Colloid In e ace Sci., 2006, 128–130, 37 and e e ences ci ed he ein. 14 Fo o he glyconano ings based on pseudo o axanes-like com- plexes using cyclodex ins see:(a) T. Ooya, M. Eguchi and N. Yui, J. Am. Chem. Soc., 2003, 125, 1140; (b) A. Nelson, J. M. Beli sky, S. Vidal, C. S. Joine , L. G. Baum and J. S odda , J. Am. Chem. Soc., 2004, 126, 11914. 15 (a) S. A. Lasky, Annu. Re . Biochem., 1995, 64, 113; (b) R. Bane jee, K. Das, R. Ra ishanka , K. Suguna, A. Su olia and A. M. Vijayan, J. Mol. Biol., 1996, 259, 281. 16 S. De and A. Su olia, J. Biosci., 2006, 31, 551. Fig. 4 TEM cha ac e iza ion o he specifici y acqui ed by he agg ega es owa d specific ecep o s. (A) In e ac ion o PNA wi h SWCNT-Lac-8. The abacus like geome y is los as he s ia ions a e co e ed by lec in molecules (whi e a ows). (B) In e ac ion o PNA wi h SWCNT-Cellob-9. The e is no specific in e ac ion and he cha ac e is ic s ia ions a e s ill obse ed. (C) TEM image o pu ified PNA lec in alone. This jou nal is cThe Royal Socie y o Chemis y 2009 Chem. Commun., 2009, 4121–4123 |4123 Published on 02 June 2009. Downloaded by G al Uni e sidad Se illa on 22/06/2016 16:23:44. View A icle Online