Nanocapsules of Magnetic Au Self-Assembly for DNA Migration and

Jan 17, 2018 - A conclusion could be obtained from the previous reports that Au VLAs are ideal self-assembly scaffolds providing abundant conjugation ...
0 downloads 9 Views 732KB Size
Subscriber access provided by READING UNIV

Article

Nanocapsules of Magnetic Au Self-Assembly for DNA Migration and Secondary Self-Assembly Ling Wang, Yitong Wang, Shuli Dong, Yongming Deng, and Jingcheng Hao ACS Appl. Mater. Interfaces, Just Accepted Manuscript • DOI: 10.1021/acsami.7b18689 • Publication Date (Web): 17 Jan 2018 Downloaded from http://pubs.acs.org on January 17, 2018

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

ACS Applied Materials & Interfaces is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

1DQRFDSVXOHV RI 0DJQHWLF $X 6HOI $VVHPEO\ IRU '1$ 0LJUDWLRQ DQG 6HFRQGDU\ 6HOI $VVHPEO\ /LQJ :DQJ D GN G FRQF @ OLQHDU FXUYHV (DFK VDPSOH ZDV PHDVXUHG WKUHH WLPHV IRU WKH DYHUDJH YDOXH $X 13V ZHUH V\QWKHVL]HG DFFRUGLQJ WR D SUHYLRXV UHSRUW +$X&O Ü + 2 P/ DW

PJ ZDV GLVVROYHG LQ D PL[WXUH RI WHWUDOLQ

7\SLFDOO\

P/ DQG 2/

ž& WR REWDLQ DQ RUDQJH VROXWLRQ XQGHU 1 IORZ 8QGHU YLJRURXV VWLUULQJ D

UHGXFLQJ DJHQW VROXWLRQ ZLWK 7%

%$PJ 2/

P/ DQG WHWUDOLQ

P/

ZHUH TXLFNO\ LQMHFWHG LQWR WKH VROXWLRQ RI +$X&O 7KH UHGXFWLRQ ZDV LQVWDQWDQHRXVO\ LQLWLDWHG DQG WKH VROXWLRQ FKDQJHG WR D GHHS SXUSOH FRORU ZLWKLQ DOORZHG WR UHDFW DW U W IRU

K XQGHU 1 IORZ $IWHU DGGLWLRQ RI

ACS Paragon Plus Environment

V 7KH PL[WXUH ZDV P/ HWKDQRO $X 13V

Page 9 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

ZHUH FROOHFWHG E\ FHQWULIXJDWLRQ KH[DQH DQG

USP

PLQ DQG ZDVKHG WZLFH ZLWK

P/

P/ HWKDQRO

7KH FDWLRQLF VXUIDFWDQW &7$)H

ZDV SUHSDUHG DV UHSRUWHG E\ PL[LQJ HTXDO PRODU

DPRXQW RI &7$% DQG )H&O Ü + 2 LQ PHWKDQRO DQG VWLUULQJ RYHU QLJKW DW URRP WHPSHUDWXUH DQG WKHQ GHK\GUDWHG LQ YDFXR DW

ƒ& RYHUQLJKW &7$&H ZDV

V\QWKHVL]HG E\ WKH VDPH PHWKRG &7$&H ZDV GLVVROYHG LQ ZDWHU DQG NHSW LQ D PLQ WR SUHSDUH D

PJ P/ VROXWLRQ $

ƒ& ZDWHU EDWK IRU DSSUR[LPDWHO\ / &7$&H VROXWLRQ DQG D

QDQRSDUWLFOH VROXWLRQ RQH WHQWK RI $X 13V SURGXFHG IURP KH[DQH ZHUH SODFHG LQWR D XOWUDVRXQG IRU

PJ +$X&O Ü + 2

P/ PLFURFHQWULIXJH WXEH ZKLFK ZDV FDSSHG DQG NHSW

PLQ 7KH PLFURFHQWULIXJH WXEH ZLWKRXW FDS ZDV WKHQ SODFHG LQWR D

JHQWO\ VWLUUHG ZDWHU EDWK DW

ƒ& IRU a

K

7KH QDQRFDSVXOHV RI $X 13V &7$&H '1$ ZHUH SUHSDUHG E\ DGGLQJ /

/ RI

PPRO / '1$ VROXWLRQ WR D

GURS RI

P/ $X 13V &7$&H 9/$ VROXWLRQ 7KH DGGHG

'1$ GURSOHWV LPPHGLDWHO\ IRUPHG IORFFXODWHV RQFH WKH GURSOHWV FRQWDFW ZLWK WKH VROXWLRQV $IWHU K LQFXEDWLRQ DW

ƒ& '1$ QDQRFDSVXOHV FDQ EH REWDLQHG

5(68/76 $1' ',6&,66,21 )DEULFDWLRQ RI 0DJQHWLF $X 9/$V ZLWK 0DJ 6XUIV $V GHSLFWHG LQ 6FKHPH F RXU LQTXLU\ EHJDQ ZLWK WKH IDEULFDWLRQ RI VFDIIROG WHPSODWHV RI PDJQHWLF $X 9/$V E\ WKH LQQRYDWLYH 0DJ 6XUIV DVVLVWHG HPXOVLRQ VHOI DVVHPEO\ RI $X 13V $X 13V ZLWK DQ DYHUDJH VL]H RI



UHSRUWHG SURFHGXUH )LJXUH

QP FDSSHG ZLWK ROH\ODPLQH ZHUH V\QWKHVL]HG WKURXJK D $FFRUGLQJ WR HPXOVLILFDWLRQ WHFKQLTXH

ACS Paragon Plus Environment

WKH UHG

ACS Applied Materials & Interfaces

KH[DQH FRQWDLQLQJ K\GURSKRELF $X 13V ZDV PL[HG ZLWK ZDWHU SKDVH FRQWDLQLQJ VXUIDFWDQW &7$&H YROXPH UDWLR LV

IROORZHG E\ XOWUDVRQLFDWLRQ JHQHUDWLQJ DQ

RLO LQ ZDWHU HPXOVLRQ 8SRQ WKH VXEVHTXHQW HYDSRUDWLRQ RI KH[DQH DW

ƒ& $X 13V

LQ RLO GURSOHWV ZHUH SDFNHG LQWR 9/$V 6FKHPH E

F

G



QP

)UHTXHQF\

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 10 of 32

'LDPHWHU QP

)LJXUH

D 6FKHPDWLF LOOXVWUDWLRQ RI $X 13V 7KH +5 7(0 LPDJH E DQG VL]H

GLVWULEXWLRQ F RI $X 13V FDSSHG ZLWK ROH\ODPLQH

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



QP LQ )LJXUH D )LJXUH 6



QP

:KHQ KH[DQH LV

HPSOR\HG IRU HPXOVLRQ VHOI DVVHPEO\ $X 13V &7$&H LV 9/$V EXW WKHUH DUH QR

ACS Paragon Plus Environment

Page 11 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

VHOI DVVHPEOLHV IRU $X 13V &7$)H )LJXUH F $X 13V &7$% DOVR VKRZHG W\SLFDO 9/$V VWUXFWXUH E\ HYDSRUDWLQJ FKORURIRUP )LJXUH 13V &7$% 9/$V



EXW WKH $X 13V LQ $X

QP LV PXFK VPDOOHU WKDQ WKDW RI $X &7$&H

QP IRU $X 13V &7$&H KH[DQH DQG )LJXUH D )LJXUH 6

G





QP IRU $X 13V &7$&H WROXHQH LQ

%DVHG RQ WKHVH H[SHULPHQWDO UHVXOWV ZH EHOLHYH WZR PDMRU

IDFWRUV FDQ DIIHFW WKH HPXOVLRQ VHOI DVVHPEO\ SURFHVV LQ WKLV ZRUN )LUVWO\ WKH PRUSKRORJLFDO FRQWURO RI $X DVVHPEOLHV FRXOG EH DFKLHYHG E\ PDQLSXODWLQJ WKH RLO ZDWHU LQWHUIDFLDO WHQVLRQ WKURXJK MXGLFLRXV VHOHFWLRQV RI VROYHQWV DQG VXUIDFWDQWV 6HFRQGO\ WKH FRXQWHULRQV LQ VXUIDFWDQWV DOVR VKRZHG JUHDW LPSDFW WR WKH PRUSKRORJ\ RI $X DVVHPEOLHV

)LJXUH

5HSUHVHQWDWLYH 7(0 LPDJHV RI HPXOVLRQ VHOI DVVHPEOHG 9/$V IURP

PJ P/ &7$&H LQ KH[DQH $ DQG D DQG WROXHQH % DQG E LPDJHV RI QDQRVWUXFWXUH IRUPHG ZLWK &7$)H LPDJH RI DVVHPEOLHV IRUPHG IURP &7

%$& DQG F 7\SLFDO 7(0

PJ P/ KH[DQH

PJ P/ FKORURIRUP

ACS Paragon Plus Environment

' DQG G 7(0

ACS Applied Materials & Interfaces 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 12 of 32

:H LOOXVWUDWHG WKH ILUVW SRLQW RI WKH IDFWRUV ZKLFK DIIHFW $X DVVHPEOLHV $ V\VWHPDWLF LQYHVWLJDWLRQ LQWR WKH LQIOXHQFH RI HPXOVLRQ VHOI DVVHPEO\ FDXVHG E\ WKH SURSHUWLHV RI VXUIDFWDQW DQG VROYHQW LV GHPRQVWUDWHG LQ WKH IROORZLQJ SDUW $Q LPSRUWDQW VWHS IRU HPXOVLILFDWLRQ LV WR WUDQVIHU WKH K\GURSKRELF $X 13V IURP RLO SKDVH LQWR WKH RLO ZDWHU LQWHUIDFH

7KLV SURFHVV LV VSRQWDQHRXV DQG GULYHQ E\ WKH VXP RI

LQWHUIDFLDO HQHUJ\ DQG YDQ GHU :DDOV LQWHUDFWLRQV WKDW DUH EHWZHHQ WKH K\GURSKRELF FKDLQV RI VXUIDFWDQW DQG ROH\ODPLQH RQ $X 13V

,Q WKLV ZRUN WKH QRQSRODU VROYHQWV

ZLWK ORZHU ZDWHU VROXELOLW\ DQG KLJKHU LQWHUIDFLDO WHQVLRQV VXFK DV KH[DQH ZZ

WR

P1 P DQG WROXHQH

ZZ

WR

î

P1 P 7DEOH 6

H[KLELWHG VWURQJHU FDSDFLW\ LQ FRQWUROOLQJ WKH PRUSKRORJ\ RI $X 13V DVVHPEOLHV E\ 0DJ 6XUIV &7$&H RU &7$)H ZKLOH PRUH SRODU FKORURIRUP LV VXLWDEOH IRU $X 13V &7$% )LJXUH

ZZ

P1 P

7KLV UHVXOW PLJKW EH FDXVHG E\ WKH GLIIHUHQFH

LQ K\GURSKLOLF K\GURSKRELF SURSHUWLHV EHWZHHQ 0DJ 6XUIV DQG &7$% ZKLFK FRXOG SURYLGH YDOXDEOH JXLGHV IRU VROYHQW VHOHFWLRQ LQ HPXOVLRQ VHOI DVVHPEO\ 7KH 0DJ 6XUIV &7$&H DQG &7$)H SRVVHVVLQJ ODUJHU FRXQWHULRQ >&H&O %U@ RU >)H&O %U@ WKDQ %U ZHUH H[SHFWHG WR EH OHVV HIIHFWLYH WR VFUHHQ WKH UHSXOVLRQV EHWZHHQ DGMDFHQW FDWLRQ FDWLRQ KHDG JURXSV DQG WR LQFUHDVH WKH FULWLFDO PLFHOOH FRQFHQWUDWLRQ FPF +RZHYHU WKH FPF YDOXHV RI &7$&H

PPRO / DQG &7$)H

PPRO /

PHDVXUHG E\ HOHFWULFDO FRQGXFWLYLW\ PHDVXUHPHQWV ZHUH ORZHU WKDQ &7$% PPRO /

)LJXUH

(DVWRH HW DO

LOOXPLQDWHG WKDW WKH LQWHUDFWLRQ EHWZHHQ

K\GURSKRELF DQLRQV LQ 0DJ 6XUIV DQG K\GURSKRELF PRLHWLHV RI DON\O FKDLQ LQFUHDVH WKH RYHUDOO K\GURSKRELF SURSHUW\

7KHUHIRUH WKH K\GURSKRELF 0DJ 6XUIV H[KLELW

VWURQJHU VHOI DVVHPEO\ FDSDELOLW\ ZLWK RUJDQLF SKDVH RI KH[DQH RU WROXHQH DQG K\GURSKLOLF &7$% SURYLGHV UREXVW HPXOVLRQ VHOI DVVHPEO\ LQ FKORURIRUP $GGLWLRQDOO\ $X 13V &7$&H VKRZHG W\SLFDO 9/$V VWUXFWXUH E\ HYDSRUDWLQJ KH[DQH

ACS Paragon Plus Environment

Page 13 of 32

EXW QRYHO GLVFRLG DVVHPEOLHV LQ WKH WROXHQH )LJXUH D DQG E 89 9LV VSHFWURVFRS\ RI $X 13V &7$&H VROXWLRQ VKRZHG DQ REYLRXV UHG VKLIW LQ WKH ORFDOL]HG VXUIDFH SODVPRQ UHVRQDQFH /635 SHDN IURP KH[DQH DQG

QP )LJXUH 6

QP HYDSRUDWLQJ WROXHQH

WR

QP HYDSRUDWLQJ

ZKLFK UHVXOWHG IURP DQ LQFUHDVLQJ

SODVPRQLF FRXSOLQJ RI DGMDFHQW $X 13V DQG VL]H LQFHQVHPHQW RI $X 13V )LJXUH 6 7KLV SURFHVV ZDV DFFRPSDQLHG E\ D FRORU FKDQJH RI WKH $X VROXWLRQ IURP ZLQH UHG WR SXUSOH )LJXUH 6

&RQWUDVW WR QRQSRODU VROYHQWV PRUH SRODU FKORURIRUP ZDV PRUH

DSSOLFDEOH IRU &7$% DVVLVWHG HPXOVLILFDWLRQ DSSURDFK SURGXFLQJ ZHOO GHILQHG VSKHULFDO DVVHPEOLHV ZLWK H[WUHPHO\ FRQGHQVHG $X 13V LQ )LJXUH

' DQG G 7KH

VROYHQW HIIHFW RQ WKH LQWHUIDFLDO WHQVLRQ DQG VHOI DVVHPEO\ SURFHVV ZHOO DFFRXQWV IRU WKH GLIIHUHQW DVVHPEO\ PRUSKRORJLHV RI $X DVVHPEOLHV WKH VROYHQW ZLWK KLJKHU LQWHUIDFLDO WHQVLRQ KH[DQH VKRXOG DWWUDFW $X 13V PRUH VWURQJO\ WKDQ WROXHQH DQG PRUH SRODU FKORURIRUP FDQQRW EH VXLWDEOH IRU 0DJ 6XUIV EXW &7$% LQ HPXOVLILFDWLRQ SURFHVV

D

E

N P6 FP

N P6 FP

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

F&7$)H PPRO /

F&7$&H PPRO /

)LJXUH

(OHFWULFDO FRQGXFWLYLW\ GDWD

N

YV

&7$&H

D

DQG &7$)H

FRQFHQWUDWLRQ 7KH EUHDN SRLQW LV WKH FULWLFDO PLFHOOH FRQFHQWUDWLRQ

FPF

E RI

VXUIDFWDQWV (DFK N ZDV WKH DYHUDJH YDOXH FDOFXODWHG IURP WKH WKUHH PHDVXUHPHQWV 7 “

ž&

ACS Paragon Plus Environment

ACS Applied Materials & Interfaces 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 14 of 32

)RU WKH VHFRQG SRLQW WKH FRXQWHULRQV LQ VXUIDFWDQWV LQFOXGLQJ >&H&O %U@ >)H&O %U@ DQG %U KDYH JUHDW LPSDFW WR WKH PRUSKRORJ\ RI $X DVVHPEOLHV 7KRXJK SURFHHGLQJ HPXOVLRQ VHOI DVVHPEO\ ZLWK D VDPH RUJDQLF SKDVH RI KH[DQH )LJXUH D DQG

F

$X 13V &7$&H SUHVHQWV FROORLGDO 9/$V EXW ZLWKRXW IRUPDWLRQ RI

VHOI DVVHPEOHG VWUXFWXUHV IRU $X 13V &7$)H VXEVWDQFHV ZLWK MXMXEH OLNH VKDSH DUH SXUH &7$)H SURYHG LQ )LJXUH 6

1RWHZRUWK\ WKH >&H&O %U@ RU >)H&O %U@ FRXOG

LQGXFH $X 13V ZLWK D VSRQWDQHRXV VL]H JURZWK SKHQRPHQRQ XQGHU HPXOVLRQ VHOI DVVHPEO\ SURFHVV KRZHYHU $X 13V NHSW WKHLU RULJLQDO VL]H LQ $X 13V &7$% $V VKRZQ LQ )LJXUHV

DQG 6

WKH FRXSOHG $X 13V LQ $X 13V &7$&H DQG $X

13V &7$)H DVVHPEOLHV VKRZHG D VL]H VHOI JURZQ SURFHVV IURP RULJLQDO WR



DQG

QP $X 13V &7$&H KH[DQH “





QP

QP $X 13V &7$&H WROXHQH

QP $X 13V &7$)H KH[DQH 7KLV REVHUYDWLRQ LV GLIIHUHQW IURP DOO

RI UHSRUWHG UHVXOWV

LQ ZKLFK 13V VXFK DV $X DQG )H 2 13V NHSW WKHLU RULJLQDO

VL]H E\ XVLQJ HPXOVLRQ VHOI DVVHPEO\ PHWKRG :H EHOLHYH WKDW WKH R[LGL]DELOLW\ RI >&H&O %U@ DQG >)H&O %U@ DUH DFFRXQW IRU WKH VHOI JURZWK SKHQRPHQRQ RI $X 13V 2XU SUHYLRXV ZRUN KDV SURYHG WKH VWURQJ R[LGL]DELOLW\ RI >)H&O %U@ LQ &7$)H PD\ OHDG WR WKH UHODWLYHO\ ODUJH $X#&7$) QDQRSDUWLFOHV a

QP DQG H[FHVV 1D%+

FRXOG EULQJ DERXW WKH UHGXFWLRQ RI >)H&O %U@í DQG WKH LQWURGXFWLRQ RI LPSXULWLHV 5HSODFLQJ &7$)H ZLWK D ZHDN R[LGL]LQJ 0DJ 6XUIV & +

&+ 1 >&H&O %U@

&7$&H RU &7$*G

IRU VXUIDFH PRGLILFDWLRQ RQ $X 13V WKH PDJQHWLF

$X#&7$&H RU $X#&7$*G QDQRSDUWLFOHV ZHUH V\QWKHVL]HG ZLWK XOWUDILQH VL]H RI QP

$FFRUGLQJ WR WKLV PHFKDQLVP IRU RXU ZRUN WKH H[FHVV UHGXFWDQW RI

WHUW EXW\ODPLQH ERUDQH FRPSOH[ 7%$% FRXOG UHVXOW LQ WKH UHGXFWLRQ RI >&H&O %U@í RU >)H&O %U@ DQG OHDG WR WKH VL]H VHOI JURZQ SURFHVV RI $X 13V %HVLGHV VWURQJHU

ACS Paragon Plus Environment

Page 15 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

R[LGL]DELOLW\ RI >)H&O %U@ WKDQ >&H&O %U@ LQGXFHG ODUJHU VL]H RI $X 13V LQ $X 13V &7$)H FRPSDUHG ZLWK WKDW RI $X 13V &7$&H LQ )LJXUH D DQG F 7KH &DSDELOLW\ RI &RQJOXWLQDWLRQ DQG &RQWUROODEOH 0LJUDWLRQ RI '1$ E\ $X 13V &7$&H 6FDIIROG 0RUHRYHU WKH SHUIHFW FRQMXJDWLRQ RI $X DQG &7$&H WKURXJK K\GURSKRELF LQWHUDFWLRQV PDNHV $X 13V &7$&H 9/$V DQ LGHDO VHOI DVVHPEO\ VFDIIROG ZLWK ZHOO GHILQHG VWUXFWXUH DYHUDJH VL]H LV ]HWD SRWHQWLDO RI



P9

QP DQG SRVLWLYH FKDUJHV

7KHUHIRUH WKHVH SRVLWLYHO\ FKDUJHG $X

13V &7$&H 9/$V FDQ VHUYH DV HIILFLHQW FDUULHUV IRU FRQJOXWLQDWLRQ RI '1$ '1$ XVHG IRU VWXG\LQJ WKH FRQJOXWLQDWLRQ DQG PLJUDWLRQ KDV

EDVH SDLU ES

,Q DGGLWLRQ

ZKHQ '1$ ZDV DGKHUHG E\ $X 13V &7$&H 9/$V GXH WR WKH UHGXFWLRQ RI FKDUJH UHSXOVLRQ EHWZHHQ WKH DGMDFHQW SKRVSKDWH JURXSV PL[WXUHV RI $X 13V &7$&H DQG '1$ ZRXOG DJJUHJDWH DQG SUHFLSLWDWH RXW IURP WKH VROXWLRQV 7KH SKDVH GLDJUDP LQ )LJXUH

D VKRZV WKDW WKH PDFURVFRSLFDO SKDVH VHSDUDWLRQ RI

RFFXUUHG DW

PRO / $X 13V &7$&H EXW DW

PRO / '1$

PRO / ZLWK SXUH &7$&H ,W

LQGLFDWHG WKH VWURQJHU '1$ ELQGLQJ FDSDELOLW\ RI $X 13V &7$&H WKDQ &7$&H $FFRUGLQJ WR 89 9LV PRQLWRULQJ )LJXUH E WKH RSWLFDO GHQVLW\ RI >

PD[

QP 2'

@ GHFUHDVHG IURP

WR

VROXWLRQ 7KHVH REVHUYDWLRQV LQGLFDWHG WKDW

LQ

PRO / '1$

PRO / $X 13V &7$&H

'1$ ZHUH DGKHUHG WR $X

13V &7$&H 9/$ ZKLFK LV D PRUH HIILFLHQW FDUULHU WKDQ &7$&H ZLWK

'1$

FRQJOXWLQDWLRQ 7KH UHVXOWV IURP &' H[SHULPHQW )LJXUH F ZHUH DOVR FRQVLVWHQW ZLWK WKH DIRUHPHQWLRQHG H[SHULPHQWDO UHVXOWV ZKLFK VXSSRUW $X 13V &7$&H H[KLELWHG VWURQJHU '1$ ELQGLQJ FDSDELOLW\ WKDQ &7$&H ,Q )LJXUH F '1$ SUHVHQWV W\SLFDO % IRUP DGVRUSWLRQ ZLWK D SRVLWLYH EDQG DW DURXQG VWDFNLQJ DV ZHOO DV D QHJDWLYH EDQG DW DURXQG

QP FRUUHVSRQGLQJ WR WKH EDVH QP EURXJKW E\ WKH KHOLFLW\

)RU

$X 13V &7$&H PRUH VLJQLILFDQW UHGXFWLRQ LQ RSWLFDO LQWHQVLW\ RI ERWK ERQGV

ACS Paragon Plus Environment

ACS Applied Materials & Interfaces

FRQWULEXWHV WR WKH FRQMXJDWLRQ RI $X DQG &7$&H ZKLFK HQDEOHV $X 13V DFW DV VHOI DVVHPEO\ VFDIIROG ORDGLQJ &7$&H ZLWK PRUH SRVLWLYH FKDUJHV WR DGKHUH '1$ UHVXOWLQJ LQ D VWURQJHU ELQGLQJ FDSDELOLW\ RI '1$ PROHFXOHV

D

&7$&H

$X &7$&H

&RQFHQWUDWLRQ PPRO /

E

F

'1$ &7$&H $X &7$&H

&' PGHJ

'1$ &7$&H $X &7$&H

$EV D X

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 16 of 32

:DYHOHQJWK QP

:DYHOHQJWK QP

)LJXUH

D 3KDVH EHKDYLRU RI

PRO / '1$ PL[HG ZLWK YDULRXV DPRXQWV RI

&7$&H DQG $X 13V &7$&H UHVSHFWLYHO\ 7ULDQJOH FRUUHVSRQGV WR FOHDU VROXWLRQV ZKHUHDV FLUFOHV FRUUHVSRQG WR WXUELG RU PDFURVFRSLFDOO\ SKDVH VHSDUDWHG VDPSOHV 89 9LV VSHFWUD E DQG &' F VKRZ WKH VHOI DVVHPEO\ EHKDYLRU RI LQ WKH SUHVHQFH RI

PRO / '1$

PRO / &7$&H RU $X 13V &7$&H

$V SUHYLRXVO\ UHSRUWHG

PDJQHWLF VXUIDFWDQW '7$&H

& +

>&H&O %U@í FRXOG IDEULFDWH IHUURIOXLGV RI WKHUPRWURSLF OLTXLG FU\VWDOV

1 &+ >&H&O %U@í

DOORZHG WKHLU PDJQHWLF SURSHUWLHV WR EH FRQWUROOHG E\ H[WHUQDO PDJQHWLF ILHOG 6XSHUFRQGXFWLQJ TXDQWXP LQWHUIHUHQFH GHYLFH PDJQHWRPHWU\ )LJXUH

D FOHDUO\

VKRZV WKH SDUDPDJQHWLF SURSHUWLHV RI &7$&H ZLWK D SDUW RI IHUURPDJQHWLF SURSHUW\ DV ZHOO DV WKH SDUDPDJQHWLF &7$)H LQ )LJXUH 6

+HUHLQ ZH SURSRVHG WKDW >&H&O %U@í

ACS Paragon Plus Environment

Page 17 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

FRXSOHG $X 13V &7$&H 9/$V VHUYLQJ DV D VHOI DVVHPEO\ VFDIIROG ZRXOG HQKDQFH WKH FRQJOXWLQDWLRQ VHQVLWLYLW\ RI '1$ DQG LQGXFH WDUJHW '1$ PLJUDWLRQ 7R SURYH WKLV K\SRWKHVLV WKH H[SHULPHQWV ZHUH SHUIRUPHG ZLWK D ZHDN 1G)H% PDJQHW SRVLWLRQHG DW WKH ERWWRP RI WKH VROXWLRQ FRPSRVLQJ

PRO / '1$ DQG

7 PRO /

$X 13V &7$&H 9/$V $X 13V &7$&H 9/$V ZLWK PDJQHWLF >&H&O %U@í FRXOG EH DWWUDFWHG WRZDUG WKH PDJQHW SURGXFLQJ DQ HQULFKHG ORFDO FRQFHQWUDWLRQ RI $X 13V &7$&H RQ WKH ERWWRP ZKLFK IXUWKHU LQGXFHG DJJUHJDWLRQ RI '1$ 8OWLPDWHO\ WKH VLJQLILFDQW UHGXFWLRQ RI '1$ LQ WKH ERWWRP ZRXOG IXUWKHU LQGXFH '1$ LQ WKH XSSHU VROXWLRQ WR PLJUDWH WRZDUGV WKH PDJQHW GHPRQVWUDWLQJ DQ REYLRXV UHGXFWLRQ LQ 2'

DV UHDFWLRQ WLPH ZHQW RQ ZLWK 89 9LV PRQLWRULQJ )LJXUH E

DERXW

$IWHU

K

'1$ PLJUDWHG WRZDUGV WKH PDJQHW ,Q FRQWUDVW ZLWKRXW PDJQHW ILHOG

)LJXUH F WKH XSSHU OD\HU RI '1$ VROXWLRQ ZDV RQO\ UHGXFHG E\

FDXVHG E\

PRO / $X 13V &7$&H VROXWLRQ 2EYLRXVO\ WKH DSSOLFDWLRQ RI PDJQHWLF ILHOG FRXOG FDXVH DQ LQFUHDVH RI

'1$ FRQJOXWLQDWLRQ E\ WKH VDPH $X 13V &7$&H

VROXWLRQ 7KH FRQWUROODELOLW\ RI WDUJHWHG '1$ PLJUDWLRQ LQGXFHG E\ H[WUD PDJQHWLF ILHOG ZDV DOVR FRQILUPHG E\ &' PHDVXUHPHQWV )LJXUH GHVFULELQJ SKHQRPHQRQ

FROORLGDO

SDUWLFOHV

SURYLGH

D

UHDVRQDEOH

G

&ODVVLFDO PRGHOV

H[SODQDWLRQ

IRU

WKLV

,Q WKH FODVVLFDO PRGHOV HOHFWULF GRXEOH OD\HUV RI FROORLGDO SDUWLFOHV

LQFOXGH 6WHUQ OD\HUV DQG GLIIXVH OD\HUV ,Q 6WHUQ OD\HU FRXQWHULRQV DUH VWURQJO\ ERXQG DQG PRYH ZLWK WKH FROORLGDO SDUWLFOHV DV D ZKROH G\QDPLF HQWLW\ &RXQWHULRQV LQ WKH GLIIXVH OD\HUV FRXOG PRYH IUHHO\ LQ WKH EXON DQG NHHS G\QDPLF HTXLOLEULXP ZLWK WKRVH LQ WKH 6WHUQ OD\HU >&H&O %U@ DUH K\GURSKRELF DQG SUHVHQW VWURQJ K\GURSKRELF LQWHUDFWLRQ ZLWK WKH DON\O FKDLQV RI &7$&H

DQG FRXOG HQWHU WKH FRUH RI $X

13V &7$&H DV D SDUW LQ 6WHUQ OD\HUV 7KHUHIRUH WKH HQULFKHG PDJQHWLF >&H&O %U@í LQ 6WHUQ OD\HU FRXOG DFW DV D ZKROH G\QDPLF HQWLW\ RI $X 13V &7$&H E\ WKH PDJQHW

ACS Paragon Plus Environment

ACS Applied Materials & Interfaces

E

$EV D X

0DJQHWLF 0RPHQW HPX J

D

0DJQHWLF )LHOG 2H

:DYHOHQJWK QP

F

G

&' PGHJ

$EV D X

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 18 of 32

PDJQHW ZLWKRXW PDJQHW

:DYHOHQJWK QP

:DYHOHQJWK QP

)LJXUH

D 648,' PDJQHWRPHWU\ RI SDUDPDJQHWLF &7$&H DW

. E 89 9LV RI

WKH UHGXFWLRQ RI '1$ RQ WKH WRS RI $X 13V &7$&H VROXWLRQ ZKHUH '1$ ZDV PRO / DQG $X 13V &7$&H 7 ILHOG

PRO / YV WLPH K WR DQ H[WHUQDO PDJQHWLF ILHOG

F 89 9LV PHDVXUHPHQW RI

PRO / '1$ ZLWK DQ H[WHUQDO PDJQHWLF

7 DQG '1$ $X 13V &7$&H

PHDVXUHPHQW ZDV SHUIRUPHG DIWHU

PRO / VROXWLRQ ZLWKRXW PDJQHW 7KH

K LQFXEDWLRQ WR VKRZ WKH UROH RI H[WHUQDO

PDJQHWLF ILHOGV DV ZHOO DV $X 13V &7$&H IRU '1$ PLJUDWLRQ G &' HYLGHQFH RI WKH UHGXFWLRQ RI '1$ RQ WKH WRS RI $X 13V &7$&H VROXWLRQ YV WLPH K WR DQ H[WHUQDO PDJQHWLF ILHOG

7

&RQVWUXFWLRQ RI $X 13V &7$&H '1$ 1DQRFDSVXOHV E\ $X 13V &7$&H 9/$V 6FDIIROG 7KH UHSRUWHG UHVHDUFKHV RI $X DVVHPEOLHV ZHUH OLPLWHG LQ WKH DSSOLFDWLRQV RI $X DVVHPEOLHV LWVHOI 7KH GHVLJQ RI QRYHO VHOI DVVHPEO\ RI VPDUW PXOWLIXQFWLRQDO K\EULG FRPSDUWPHQWV ZLWK $X 13V DVVHPEO\ LV LQ JUHDW GHPDQG IRU H[SHQGLQJ WKHLU DSSOLFDWLRQ ILHOG 6HTXHQWLDOO\ ZH UHSRUWHG KHUH D FRQYHQLHQW DQG HIILFLHQW PHWKRG WR

ACS Paragon Plus Environment

Page 19 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

HQJLQHHU WKH QRYHO '1$ QDQR DVVHPEOLHV ZLWK UHVSRQVLYHQHVV DQG IXQFWLRQDOLW\ E\ ZUDSSLQJ $X 13V &7$&H VFDIIROG ZLWK '1$ RXWVLGH $V GHVFULEHG LQ )LJXUH D GURSOHWV RI KLJKO\ FRQFHQWUDWHG '1$

ES

PPRO / VROXWLRQ ZDV DGGHG LQWR

$X 13V &7$&H VROXWLRQ VPRRWK IDFHG '1$ QDQRFDSVXOHV ZLWK KROORZ VWUXFWXUH )LJXUH E FRXOG EH SURGXFHG DIWHU

K LQFXEDWLRQ DW

ƒ& (OHPHQW PDSSLQJ LPDJH

DQG ('6 DQDO\VLV LQ )LJXUH 6 FOHDUO\ VKRZHG WKH $X HOHPHQW LQ WKH $X 13V &7$&H VFDIIROG DQG 3 HOHPHQW E\ FRDWLQJ '1$ PROHFXOHV LQ QDQRFDSVXOHV )LJXUH E $X 13V &7$&H 9/$V SURYLGHG VFDIIROG WHPSODWHV DV GDUNHU FRUH DUHD DQG '1$ ZUDSSLQJ DURXQG ZDV UHSUHVHQWHG E\ WKH OLJKWHU VKHOO DUHD )LJXUH F '1$ ZUDSSHUV VXUURXQGLQJ $X 13V LQ WKH QDQRFDSVXOHV UHGXFHG WKH LQWHUVWLWLDO VSDFH DQG YLVLELOLW\ RI $X 13V

)LJXUH

D

6FKHPDWLF GLDJUDP LOOXVWUDWLQJ WKH SUHSDULQJ PHWKRG RI $X

13V &7$&H '1$ QDQRFDSVXOHV

6(0

E

DQG 7(0

ACS Paragon Plus Environment

F

LPDJHV RI $X

ACS Applied Materials & Interfaces 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

13V &7$&H '1$

ES

Page 20 of 32

PPRO / '1$ FDSVXOHV G 3KDVH EHKDYLRU IRU $X

13V &7$&H ZLWK LQFUHDVLQJ FRQFHQWUDWLRQ RI '1$

ES 2WKHU HOHPHQWV ZHUH

FRQVLGHUHG WR VWXG\ WKH IDEULFDWLRQ RI $X 13V &7$&H '1$ FDSVXOHV LQFOXGLQJ WKH FRQFHQWUDWLRQ DQG ES QXPEHU RI '1$ PROHFXOHV DQG WKH VFDIIROG IRUPHG E\ $X H 6(0 UHVXOW RI $X 13V &7$&H '1$

ES

VKRZV WKH PRUSKRORJ\ RI $X 13V &7$&H '1$ 7(0 LPDJH RI &7$&H '1$ LPDJHV RI $X 13V &7$% '1$

ES ES

PPRO / SUHFLSLWDWH I 6(0 LPDJH ES

PPRO / SUHFLSLWDWH J

PPRO / SUHFLSLWDWH ZLWKRXW $X K 6(0 PPRO / FDSVXOHV

7KH IRUPDWLRQ RI WKHVH $X 13V &7$&H '1$ QDQRFDSVXOHV GLIIHUV IURP WKH SUHFLSLWDWH REWDLQHG IURP WKH GLOXWHG '1$ VROXWLRQ DQG FRPSDFWLRQ DJHQWV &RPSDFWLRQ DJHQWV ZLWK SRVLWLYH PXOWLYDOHQW FKDUJHV VXFK DV FDWLRQLF VXUIDFWDQWV FRXOG FRPSDFW '1$ E\ UHOLHYLQJ WKH FKDUJH UHSXOVLRQ EHWZHHQ WKH DGMDFHQW SKRVSKDWH JURXSV RI '1$ JHQHUDWLQJ WKH DJJUHJDWH DQG SUHFLSLWDWH RI '1$ FDWLRQLF VXUIDFWDQWV FRPSOH[HV

7KH SKDVH GLDJUDP GHPRQVWUDWHG WKDW $X 13V &7$&H '1$

QDQRFDSVXOHV FRXOG RQO\ EH UHFHLYHG E\ WKH DGGLWLRQ RI GURSOHWV ZLWK RYHU '1$

PPRO /

ES LQ )LJXUH G RWKHUZLVH SUHFLSLWDWH VLPLODU WR '1$ FDWLRQLF VXUIDFWDQW

FRPSOH[HV ZDV REWDLQHG 7KH VHOI DVVHPEO\ RI

PPRO / '1$

ES ZLWK $X

13V &7$&H VFDIIROG FRXOG DOVR SURGXFH SUHFLSLWDWH ZLWK LUUHJXODU PRUSKRORJ\ LQ )LJXUH H 7KLV UHVXOW LOOXVWUDWHV WKDW WKH FRQFHQWUDWLRQ RI '1$ PROHFXOHV PLJKW SOD\ D FULWLFDO UROH LQ WKH KLHUDUFKLFDO VHOI DVVHPEO\ SURFHVV DQG $X 13V &7$&H '1$ IDEULFDWLRQ 7KH UHSRUWHG '1$ JHO SDUWLFOHV IRUPHG E\ KLJK YLVFRVLW\ VROXWLRQ RI '1$ DQG VXUIDFWDQW &7$% FRXOG VKHG D OLJKW RQ WKH IRUPDWLRQ RI $X 13V &7$&H '1$ QDQRFDSVXOHV +LJKO\ FRQFHQWUDWHG DTXHRXV PL[WXUHV RI RSSRVLWHO\

ACS Paragon Plus Environment

Page 21 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

FKDUJHG &7$% DQG '1$ PROHFXOHV XQGHUJR DVVRFLDWLYH HOHFWURVWDWLF LQWHUDFWLRQ FRDFHUYDWLRQ DQG JHODWLRQ SURFHVV LQ ZKLFK KLJK YLVFRVLW\ VROXWLRQ RI '1$ HYRNHG WKH LQWHUIDFLDO GLIIXVLRQ SURFHVV ZKLFK FRQWULEXWHV WR WKH IRUPDWLRQ RI '1$ JHO SDUWLFOHV

:H EHOLHYH WKH LQWHUIDFLDO GLIIXVLRQ SURFHVV DURXVHG IURP KLJK YLVFRVLW\

VROXWLRQ RI '1$ VROXWLRQ !

PPRO /

ES LV WKH PDLQ UHDVRQ IRU WKH IRUPDWLRQ

RI $X 13V &7$&H '1$ QDQRFDSVXOHV UDWKHU WKDQ SUHFLSLWDWH )RU IXUWKHU SURYLQJ WKLV K\SRWKHVLV D VWUDLJKWIRUZDUG FRQWURO RYHU WKH YLVFRVLW\ RI '1$ E\ FKDQJLQJ WKHLU ES QXPEHU ZDV LQYHVWLJDWHG &KDQJLQJ '1$ ZLWK NHHSLQJ FRQVWDQW FRQFHQWUDWLRQ RI

ES LQWR WKDW FRQWDLQLQJ

ES

PPRO / DOVR SURGXFHG SUHFLSLWDWH LQ )LJXUH I

GHPRQVWUDWLQJ WKDW WKH UHGXFHG YLVFRVLW\ RI '1$ VROXWLRQ FRXOG QRW IDFLOLWDWH IRUPLQJ $X 13V &7$&H '1$ QDQRFDSVXOHV 7KH GLIIHUHQFH LQ ES QXPEHU DQG FRQFHQWUDWLRQ IRU '1$ PROHFXOHV SRVVLEO\ DFFRXQW IRU WKH YLVFRXV GHJUHH RI '1$ LQGXFLQJ FRPSOHWHO\ GLIIHUHQW LQWHUIDFLDO GLIIXVLRQ SURFHVV DQG IXUWKHU JHQHUDWLQJ D YHUVDWLOH VFHQDULR RI VHOI DVVHPEO\ LQWR QDQRFDSVXOHV RU SUHFLSLWDWH ,Q RUGHU WR REWDLQ PRUH LQVLJKW LQWR WKH $X 13V &7$&H '1$ QDQRFDSVXOHV WKH $X 13V &7$&H VFDIIROG LQFOXGLQJ $X SDUWLFOHV DQG 0DJ 6XUIV &7$&H KDV EHHQ LQYHVWLJDWHG LQ GHWDLOV %HVLGHV WKH KLJK YLVFRXV '1$ VROXWLRQ WKH H[LVWHQFH RI $X LV HVVHQWLDO IRU SUHSDULQJ '1$ QDQRFDSVXOHV LQ FRPSDULVRQ ZLWK &7$&H '1$

ES

PPRO / ZLWKRXW $X RQO\ LUUHJXODU DJJUHJDWLRQ FRXOG EH REWDLQHG LQ )LJXUH J 7KLV UHIOHFWV WKH $X 13V &7$&H VFDIIROG VKRXOG EH D NH\ HOHPHQW WR LQIOXHQFH WKH QDQRFDSVXOH FRQVWUXFWLRQ LQ DGGLWLRQ WKH VFDIIROG $X DVVHPEOLHV KDYH ZHOO FRQWURO RYHU WKH PRUSKRORJ\ DQG VWUXFWXUDO VWDELOLW\ IRU '1$ QDQRFDSVXOHV 'LIIHUHQW IURP

ACS Paragon Plus Environment

ACS Applied Materials & Interfaces 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

WKH

VWDEOH

DQG

13V &7$% '1$

VPRRWK IDFHG ES

$X

13V &7$&H '1$

Page 22 of 32

QDQRFDSVXOHV

$X

PPRO / DVVHPEOLHV SUHVHQWHG EOXUU\ DQG FROODSVHG

PRUSKRORJ\ DV VKRZQ LQ )LJXUH K 9DULDEOH VL]HV RI $X 13V LQ $X 13V &7$&H DQG $X 13V &7$% LQGXFHG D FRPSOHWHO\ GLIIHUHQW VWUXFWXUDO ULJLGLW\ WR WKHLU VFDIIROG WHPSODWH UHVXOWLQJ LQ WKH FRUUHVSRQGLQJ '1$ QDQRFDSVXOHV ZLWK YDULHG PRUSKRORJLHV DQG VWDELOLWLHV $V DIRUHPHQWLRQHG LQ )LJXUH

D VHOI JURZWK SURFHVV RI $X 13V

RFFXUUHG ZKHQ WKH\ VHOI DVVHPEOHG ZLWK 0DJ 6XUIV &7$&H RU &7$)H ZKLOH $X 13V NHSW WKH LQLWLDO V\QWKHVL]HG VL]H

QP LQ $X 13V &7$% DVVHPEOLHV 1RWDEO\

WKH VHOI JURZQ $X 13V ERRVWHG WKH VWUXFWXUDO ULJLGLW\ RI $X 13V &7$&H VFDIIROG DQG WKH VWDELOLW\ RI $X 13V &7$&H '1$ QDQRFDSVXOHV 0RUH LPSRUWDQWO\ 0DJ 6XUIV &7$&H LPSDUWHG $X 13V &7$&H VFDIIROG ZLWK VWURQJHU '1$ ELQGLQJ FDSDELOLW\ WKDQ $X 13V &7$% DVVHPEOLHV $V )LJXUH D VKRZV D SRVLWLYH EDQG DW DURXQG

QP DQG D QHJDWLYH EDQG DW

EDVH VWDFNLQJ DQG KHOLFLW\ RI '1$ UHVSHFWLYHO\

QP DUH DVFULEHG WR WKH

7KLV SURYHV WKH H[LVWHQFH RI

UHVLGXDO '1$ PROHFXOHV LQ WKH VXSHUQDWDQW RI $X 13V &7$% '1$ DVVHPEOLHV DIWHU FHQWULIXJDWLRQ

ZKLOH QR '1$ PROHFXOHV FRXOG EH GHWHFWHG LQ WKDW RI $X

13V &7$&H '1$ QDQRFDSVXOHV 7KH VLPLODU FRQFOXVLRQ FRXOG EH REWDLQHG IURP )LJXUH E D /635 SHDN DW

QP DOVR FRQILUPHG WKH SUHVHQFH RI $X 13V LQ WKH

VXSHUQDWDQW LQ $X 13V &7$% '1$ DVVHPEOLHV 7KH VWURQJHU '1$ ELQGLQJ FDSDELOLW\ IRU $X 13V &7$&H VFDIIROG LQGXFHG E\ 0DJ 6XUI &7$&H PLJKW EH DQRWKHU UHDVRQ IDFLOLWDWLQJ $X 13V &7$&H '1$ IDEULFDWLRQ $ V\VWHPDWLF ZRUN FRQFHUQLQJ WKH '1$ FRPSDFWLRQ GLVFRYHUHG WKDW FDWLRQLF VXUIDFWDQW FHW\OWULPHWK\ODPPRQLXP ZLWK

ACS Paragon Plus Environment

Page 23 of 32

%U &7$% DQG &O &7$&O DV FRXQWHULRQV FRXOG FDXVH D GHFRPSDFWLRQ EHKDYLRU RI '1$

%XW FDWLRQLF VXUIDFWDQW ZLWK PDJQHWLF >)H&O %U@ &7$)H KDV DQ DPD]LQJ

'1$ FRQGHQVH FDSDELOLW\ HYHQ DW DQ H[WUHPHO\ KLJK FRQFHQWUDWLRQ

7KH\ EHOLHYH

WKDW WKH VWURQJ K\GURSKRELFLW\ RI PDJQHWLF >)H&O %U@ PDNHV LW FDSDEOH WR HQWHU WKH FRUH RI PLFHOOHV E\ K\GURSKRELF LQWHUDFWLRQ ZLWK DON\O FKDLQV

SURGXFLQJ PRUH

DFWLYH VLWHV IRU FDWLRQLF KHDG JURXSV WR LQWHUDFW ZLWK '1$ UHVXOWLQJ LQ D VXVWDLQHG '1$ FRPSDFWLRQ FDSDELOLW\ DW KLJK FRQFHQWUDWLRQ

:H VXJJHVW WKDW WKH VWURQJ

K\GURSKRELFLW\ RI >&H&O %U@ DFFRXQW IRU WKH VWURQJHU '1$ ELQGLQJ FDSDELOLW\ IURP $X 13V &7$&H WKDQ $X 13V &7$% DVVHPEOLHV 7KHUHIRUH D FRPSUHKHQVLYH LQVLJKW LQWR WKH IRUPDWLRQ RI $X 13V &7$&H '1$ QDQRFDSVXOHV LV GHHSO\ LOOXVWUDWHG IRU IRXU SRLQWV WKH LQWHUIDFWLDO GLIIXVLRQ IURP KLJK YLVFRXV '1$ VROXWLRQ VFDIIROG $X WHPSODWH &7$&H LPSDUWHG VWUXFWXUDO ULJLGLW\ WR $X 13V &7$&H DQG WKH VWURQJHU '1$ ELQGLQJ FDSDELOLW\ E\ PDJQHWLF FRXQWHULRQ >&H&O %U@ HIIHFW

D

E

$X &7$% $X &7$&H

:DYHOHQJWK QP

)LJXUH

$X &7$% $X &7$&H

$EV D X

&' PGHJ

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

:DYHOHQJWK QP

&' D DQG 89 9LV E RI WKH GLOXWHG VXSHUQDWDQW IURP FHQWULIXJLQJ $X

13V &7$&H '1$ DQG $X 13V &7$% '1$ VROXWLRQ 7KH PHDVXUHG VROXWLRQ LV GLOXWHG WR

IROG YROXPH IRU &' DQG 89 9LV VWXG\

&21&/86,216 ACS Paragon Plus Environment

ACS Applied Materials & Interfaces 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 24 of 32

,Q WKLV ZRUN ZH VXFFHVVIXOO\ HQJLQHHUHG WKH PDJQHWLF $X 13V &7$&H DVVHPEOLHV XWLOL]LQJ HPXOVLRQ VHOI DVVHPEO\ DVVLVWHG ZLWK PDJQHWLF VXUIDFWDQWV IRU WKH ILUVW WLPH 7KH FRXQWHULRQV RI VXUIDFWDQW DQG VROYHQW DUH WKH SLYRWDO IDFWRUV FRQWULEXWH WR WKH PRUSKRORJLFDO FRQWURO RI $X DVVHPEOLHV

,Q SDUWLFXODU

PDJQHWLF FRXQWHULRQ

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v $662&,$7(' &217(17V 6XSSRUWLQJ ,QIRUPDWLRQ 7KLV LQIRUPDWLRQ LV DYDLODEOH IUHH LQ WKH FKDUJH YLD WKH LQWHUQHW DW KWWS SXEV DFV RUJ

ACS Paragon Plus Environment

Page 25 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

ACS Applied Materials & Interfaces

3URSHUWLHV RI RUJDQLF VROYHQWV VL]H GLVWULEXWLRQ DQDO\VLV RI $X 13V LQ DVVHPEOLHV 89 9LV VSHFWUXP RI $X 13V LQ KH[DQH DQG DVVHPEO\ $X 13V &7$&H VWUXFWXUHV LQ WROXHQH DQG KH[DQH SKRWRJUDSKV RI $X 13V LQ KH[DQH DQG DVVHPEO\ $X 13V &7$&H 9/$V LQ ZDWHU 7(0 LPDJH RI SXUH &7$)H ZLWK WKH MXMXEH OLNH VKDSH 648,' PDJQHWRPHWU\ RI SDUDPDJQHWLF RI &7$)H DW

. 6(0 HOHPHQW PDSSLQJ LPDJH RI

$X 13V &7$&H '1$ FDSVXOHV ('6 UHVXOWV RI $X 13V &7$&H '1$ QDQRFDSVXOHV 3') v $87+25 ,1)250$7,21 &RUUHVSRQGLQJ $XWKRU Ž( PDLO MKDR#VGX HGX FQ 7HO

)D[

25&,' -LQJFKHQJ +DR 6KXOL 'RQJ