yue 2009 nano channel and its application in analytical chemistry bio-chem-12

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Nanochannel and Its Application in Analytical Chemistry

Zenglian Yue, Guoqing Zhao, Bin Peng, Shasheng Huang*

College of Life and Environment & science

Shanghai Normal University

Shanghai, 200234, China

[email protected] http://www.shnu.edu.cn

Abstract: The nanochannels method for the separation and detection of analytes plays an important role in the

analytical chemistry and is exhibiting the great potential advantages and promising future. In this review we bring

together and discuss a number of nanochannels made of biological and special material. The preparation and appli-

cation of biological nanochannels are described. Compared with the biological single channel, nanochannels pre-pared by special material are more selective and have advantages in practical application because these channels

are less fragile and more easily modified. Studies of the special material membrane involve in the chemical, bio-

technical, medical fields, etc. For the increasing interest in using material channels at present, we demonstrated the

synthetic methods of different special material nanochannels and a mount of their applications in analytical chem-

istry here. The advantages and tendency of nanochannels are discussed as well.

Key-Words: - Nanochannels, separation, preparation, applications

1 Introduction

Traditional separation methods and techniques such as

extraction, ion-exchange, chromatography, etc. are

confronted with great challenges. How to separate and

analyze trance analytes is the focus of the challenges.With the fast development of nanotechnology in 1999s,

new thoughts and opportunities have appeared for deep

research of separation. Nanochannels are the pores or

channels with diameter from 0.1 to 100 nanometers. As

an important part of nanoscience, nanochannels tech-

nique has become a new growth point. It is exhibiting

the great potential advantages and promising future

due to the size effect, chemical and physical character-

istics.

At present, the studies about nanochannels mainly

refer to the biological nanochannels and materialnanochannels. The biological nanochannels such as

bacterial -hemolysin (-HL) a heptameric protein that

spontaneously assembles in a lipid membrane has pro-

ven in the separation of peptide [1]. A single channel

(typicallyan -hemolysin channel) also can be used as

the sensing element. These sensors can detect individ-

ual analyte with channel interactions and convert these

stochastic events into a current pulse. Sutherland et al

[2] demonstrated that peptide transport through

nanopores can also be used to analyze the structure of

peptides. Depending on this device, the change in cur-

rent can be used to determine size and the concentra-

tion of the analytical species.

However, lower stability and reproducibility of tra-

ditional bilayer membrane limited its practical applica-

tion. In contrast with the biological single channel,material nanochannels have advantages in practical

application, special because these channels are less

fragile.

The studies on the special material membrane have

received intensive interest since Martin et al. reported

gold nanochannels membrane for the separation of

small molecules on the basis of molecular size [3].

Generally, material nanochannels are prepared by spe-

cial materials such as carbon nanochannels, silicon

nanochannels, nanochannel array (eg. porous alumina

membranes and the polycarbonate tracketched me-

branes) and so on. The templates of material nano-

channels include membrane template, emulsion / mi-

cromulsion template and other kinds of templates.

In this article, we will first introduce the general

configuration and the preparation of nanochannels. The

unique properties of gold nanochannels for chemistry

and bioseparation have been summarized. At the same

time, some successful examples of nanochanneles

membrane that have been applied to chemical and

biomolecular species have been introduced. The novel

bioseparation method based on nanotubules will also

RECENT ADVANCES in BIOLOGY, BIOPHYSICS, BIOENGINEERING and COMPUTATIONAL CHEMISTRY

ISSN: 1790-5125 80 ISBN: 978-960-474-141-0


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be described. The objective of this review is to intro-

duce the extremely valuable nanochannels to the bi-

ologists and chemists and encourage them to consider

nanotubules in their research.

2. Biological nanochannels

2.1 Structure and characteristics of biological

nanochannels

Biological nanochannels were first posted in 1999.

One of the typical channels is a single -hemolysin in

cross-section embedded in a lipid bilayer with a di-

ameter about 1.5-2.6 nm [4]. Ions, water and other

small molecules can go through this channel .So it can

be used as an ion-channel. The structure of-HL is

shown as figure 1

Figure 1

The important structure features of -HL are the

mouth of the channel (2.6 nm), which leads into a lar-

ger vestibule, and the stem of the channel containing a

pore with an interior diameter of 2.2 nm. The opening

between the vestibule and stem forms the limiting ap-

erture of 1.5 nm diameter, so it can only allow the sin-

gle strand DNA to go through the channel while thedouble strand DNA can not. -HL is composed of a

ring of 14 alternating lysine and glutamate side chain.

So the channel is hydrophilic in the interior while lyo-

phobic in the exterior [5].

2.2 Applications of biological nanochannels

The biophysical characteristics of poly-nucleic acids

through the channel were first researched by

Kasianowicz and his group [6]. They found that the

single molecules of DNA or RNA can be detected as

they are driven through the -hemolysin channel by

applying an electric field. During translocation, the

spread single DNA or RNA can block the channel and

transiently block the ion current, resulting in a down-

ward current pulse. The duration of the current pulses

is proportional to the length of the polymer molecules.

It has proved that this nanochannel can be used to find

differing pyridine and purine quickly [4].The charac-

teristics of DNA hairpin within millisecond can be ana-

lyzed by combining the nanochannels and the support

vector machine [7].

The interior of the -HL pore can be modified bychanging the order of amino acids. The inner geometry

and the chemical or static characteristics of the nano-

tubules were changed due to modification. Some spe-

cial analytes (e.g nucleic acid) can transport more sen-

sitively through the nanotubules membrane becausethe functional group can be modified onto the nano-

tubules of membrane using the covalence bonding and

entrapment methods. Howorka et al. bonded DNA-SH

to the inner pore to detect the complementary DNA.

When the isonucleoside was modified in the inner pore,

the current would be decreased by 30% [8,9]. Szabo et

al. covalenced biotin molecule with the 3.4kDa poly-

ethyleneglycol of monomer cysteine -106 radical and

then bonded with the deep area of -HL pore. After

modification, the ionic current would be lower by 15%

[10].

However, the stability of traditional bilayer mem-

brane is not so high and this membrane can not play

important role in practice. So, there is much room to

improve in acquiring the carrier with more channel

selectivity and higher stability. It has been proved that

solid supported object as the carrier will be more stable

than the single channel. Cornel et al. stabled the rami-

cidin on the Au surface as a biosensor and studied the

transport of electrolyte [11]. Stora et al. studied the

bacterial multipore channel on the similar surface, and

found that the R residue can close the channel[12].

3 Nanochannels of special materialNanochannels of special material are nanochannel ar-

rays prepared by the template -synthesized method.

This kind of special material nanochannels is more

selective and producible due to the flexible nanochan-

nels membrane. The application of these nanochannels

membrane was expanded due to the modification

within the pores. The templates, preparation and ap-

plication of these channels in the below were shown in

table 1.

Table 1

3.1 The templates of nanochannels

Generally, the membranes such as porous alumina

membranes and the track-etched membranes generally

can be used as template to make nanochannels. The

track-etched membranes prepared by polycarbonate,

polyacetate or other polymer material membranes are

broadly used. At present, it is a general method to syn-

thesize nanomaterials within the pores of porous alu-


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position, by this method it is possible to control the

inner diameter by changing the deposition time[24].

Nishizawa et al. synthesized many conductive poly-

mers and prepare the nanochannel or nanowire by con-trolling the deposition time[25].

Chemical polymerization is performed by immers-

ing the membrane template into the solution including

polymeric monomer and polymeric reagent. The inner

diameter can be controlled by changing the polyreac-

tion time. The outside diameter is decided by the di-

ameter of template pore. Parthasarathy et al. produced

polyacrylonitrile nanochannel by immersing the porous

alumina membranes into the solution containing acry-

lonitrile momomer[26].

Sol-gel chemistry has recently been evolved into ageneral and powerful approach for producing inorganic

materials [27, 28]. This method typically entails hy-

drolysis of a solution of a precursor molecule to obtaina suspension of colloidal particles (the sol) and then a

gel composed of aggregated sol particles. The gel is

then thermally treated to yield the desired material. It

occurred to us that sol-gel chemistry could be done

within the pores of the nanoporous template mem-

branes to obtain tubules and fibrils of a variety of in-

organic materials. Martin used this method to make

tubules and fibrils composed of polymers, metals,

semiconductors, carbon, and Li ion intercalation mate-rials[29-31]. Lakshmi et al. combined the sol-gel and

template methods to prepare fibrils and tubules of a

variety of inorganic semiconducting materials includ-

ing ZnO, WO3 and TiO2 using alumina membrane as a

template. They found that single-crystal anatase-phase

TiO2 nanostructures can be obtained via this approach

and that these nanostructures can be used as efficient

photocatalysts [32]. Like other template synthesis

methods, changing the immersion time can get the

nanochannels or nanofibrils.

Chemical vapor deposition (CVD) is a way to de-

posit the material onto the template in the vapor. The problem is that the material may block the pores of

surface and cannot deposit inside because of the fast

deposition speed. Li et al. developed a method for

producing pure carbon nanochannel fibers which in-

volved direct spinning of continuous fibers from an

aerogel of carbon nanochannels formed by CVD [33].

This process was realized through the appropriate

choice of reactants, control of the reaction conditions

and continuous withdrawal of the product with a rotat-

ing spindle used in various geometries.

3.3 Applications of special material nanochan-

nels

3.3.1 Separation of the small moleculeThe nanochannels can be acted as the molecular siev-

ing and the filtration. In 1995, Menon and Martin first

reported a commercially available microporous poly-

carbonate filter with cylindrical nanoscopic pores. The

gold nanotubules are prepared via electroless deposi-

tion of Au onto the pore walls; i.e., the pores act as

templates for the nanotubules[24]. By controlling the

Au deposition time, Au nanotubules that have effective

inside diameters of molecular dimensions (


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(Au nanochannel membrane). Water and electrolyte are

forbidden from entering these very hydrophobic

pores/nanochannels. The transition to the on state

was induced by partitioning a hydrophobic ionic spe-cies (e.g., a drug or a surfactant) into the membrane.

The membrane switches to the on state because at a

sufficiently high concentration of this ionic analyte

species, the pores/nanochannels flood with water and

electrolyte. A pH-responsive membrane was also pre-

pared by attaching a hydrophobic alkyl carboxylic acid

silane to the alumina membrane. Steinle et al.investi-

gated the transport of amiodarone, amitriptyline andbupivacaine in the hydrophobic channels [38].

3.3.3 Separation of protein and DNA

Huang et al. prepared the gold nanotubule with about55 nm of diameter by chemical deposition of gold on

polycarbonate templates membrane. After being modi-

fied by cysteine and guanide thiocyanate, thenanomembranes were studied with BSA and IgG as the

model molecules. The results showed that guanide

thiocyanate facilitated transporting of BSA by 30 to 50

times because of hydrophilic and denaturalization ef-

fects whereas IgG almost retained its transporting

speed, indicating that gold nanotubule membranes had

a good separating capability for protein[39]. The bo-

vine IgG modified onto the pores of nanomembrane

greatly impacts the transport of detecting antibodythrough the Au nanotubules. There was a larger differ-

ence of transport rate between goat anti-bovine IgG

and goat anti-cat IgG through the B-IgG-Au-Mem.

Therefore, goat anti-bovine IgG and goat anti-cat IgG

can permeate through the nanotubules membrane se-

lectively as shown in Figure2 [40]. DNA and proteins

also were reported [41, 42]. Kohli et al. prepared gold

nanochannels with inside diameters of 12 nanometers.

A "transporter", DNA-hairpin molecule, was attached

to the inside walls of these nanochannels. These

DNA-functionalized nanochannel membranes selec-

tively recognize and transport the DNA strand that iscomplementary to the transporter strand. Under opti-

mal conditions, single-base mismatch transport selec-

tivity can be obtained [43]. The nanochannels with

uniform pores can be prepared by the template of po-

rous alumina membranes. These nanomaterials include

metal, metal alloy, metal of oxide, semiconductor, and

polymerso on [44-49]. This template is also used to

prepare DNA and protein nanochannels and then detect

them [50-51].

Figure2

3.3.4 Separation of chiral drug

Synthetic bio-nanochannel membranes were developed

and used to separate two enantiomers of a chiral drug.

Lee et al. used alumina films that had cylindrical pores

with monodisperse nanoscopic diameters (for example,

20 nanometers). Silica nanochannels were chemically

synthesized within the pores of these films, and an an-

tibody selectively bindings one of the enantiomers of

the drug was attached to the inner walls of the silica

nanochannels. These membranes selectively transport

the enantiomer specifically binded to the antibody. The

enantiomeric selectivity coefficient increases as the

inside diameter of the silica nanochannels decreases

[52].

3.3.5 Separation based on different chargeBecause Au nano template membranes (Au-NTMs) are

electronically conductive, they can be charged electro-

statically in an electrolyte solution. The inner walls of

the metal tubules can be charged by modification. The

membranes reject ions with the same sign and trans-

port ions of the opposite sign. Because the sign of the

excess charge on the tubule can be changed potentio-

statically, a metal nanotubule membrane can be either

cation selective or anion selective depending on the

potential applied to the membrane [25, 53].

In addition to transmembrane potential and nano-channel diameter, solution pH value plays an important

role in determining the transport selectivity. This is

because pH determines the net charge on the protein

molecule and this, in turn, determines the importance

of the electrophoretic transport term. Yu et al. investi-

gated the transport properties of four proteins (ly-

sozyme, bovine serum albumin, carbonic anhydrase

and bovine hemoglobulins) with different sizes and pI

values[54].

3.3.6 Sensors based on the nanochannels

The nanochannel membranes can also be used as sen-sors. Martin and his coauthors have shown that elec-

trically conductive polymers with fibrillar supermo-

lecular structures can be prepared by synthesizing the

polymer within the pores of a microporous membrane.

They compared charge transport rates in fibrillar

polypyrrole with the corresponding rates in conven-

tional polypyrrole films. The results showed that the

charge transport rates in the fibrillar versions can be

significantly higher[55].

Martin et al. described an electroless deposition pro-

cedure for filling the pores in nanoporous filtration


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membranes with metal (gold) nanowires. This method

prepared ensembles of gold nanodisk electrodes in

which the nanodisks have diameters as small as 10 nm.

Cyclic voltammetric detection limits for electroactivespecies at ensembles containing 10-nm-diameter gold

disks can be as much 3 orders of magnitude lower than

those at large-diameter gold disk electrodes26. Moretto

et al. described the construction and characterization of

an electrochemical nitrate biosensor based on the ul-

trathin-film composite membrane concept. This film

separated the analyte solution from an internal sensing

solution which contained the enzyme nitrate reductaseand an electrocatalyst (methyl viologen). The sensor

showed good sensitivity to nitrate, with a detection

limit of 5.4 M and a dynamic range which extended

up to 100M NO3-

[56]. Brunetti et al prepared goldnanoelectrode ensembles (NEEs) used for biosensors

based on reductase enzymes two phenothiazines

(Azure A and B) and methylviologen. Compared with

macro electrode, NEE can obtained lower detection

limits without adsorption of the reduced forms to the

electrode surface .And it is the first use of the NEEs

for the determination of standard heterogeneous rates

constants[57].

A new kind of nanochannels membrane with coni-

cally shaped pores was developed based on the mem-

branes with cylindrical pores. The conically pores

membranes can provide dramatically higher rates oftransport than analogous cylindrical pore membranes.

Apel et al. showed that conical nanopores can be

chemically etched into radiation-tracked polymeric

membranes[58]. Li et al. investigated plasma etching

the surface of a track-etched (vide infra) polymeric

membrane (schema1) to obtain conical pores as shown

in Fig.3[33].

Figure 3

Martin research group described resistive-pulsesensing of two large DNAs, a single-stranded phage

DNA (7250 bases) and a double-stranded plasmid

DNA (6600 base pairs), using a conically shaped

nanopore in a track-etched polycarbonate membrane as

the sensing element. The conically shaped nanopore

had a small-diameter (tip) opening of 40 nm and a

large-diameter (base) opening of 1500nm. The phageDNA was driven electrophoretically through the

nanopore (from tip to base), and these translocation

events were observed as transient blocks in the ion

current. They found that the frequency of these cur-

rent-block events scales linearly with the concentration

of the DNA and with the magnitude of the applied

transmembrane potential. Increasing the applied

transmembrane potential also led to a decrease in theduration of the current-block events[59].

Sexton et al. added a protein analyte to the solution

containing antibody that selectively binds the protein.

The complex formed upon binding of the Fab to BSA

is larger than the free BSA molecule. So the cur-

rent-pulse signature for the BSA/Fab complex can be

easily distinguished from the free BSA. Furthermore,

the BSA/Fab pulses can be easily distinguished from

the pulses obtained for the free Fab and from pulses

obtained for a control protein that does not bind to the

Fab. The current-pulse signature for the BSA/Fab

complex can provide information about the size and

stoichiometry of the complex[60].

4 Conclusions

Nanotubules technology is exhibiting the great poten-

tial advantages and promising future due to the sizeeffect and chemical physical characteristics. The ap-

plications of nanochannels in many fields, such as

analytical chemistry, biochemistry, materials science,

environmental science, and so on, will undergo more

development although some reports on the nanochan-

nels have been published.

Acknowledgment

This work was supported by the National High-tech

R&D program (863 program, 2007AA06Z402), Pro-

ject of Shanghai Municipal Government

(08520510400), Shanghai Leading Academic Disci-

pline Project (S30406), and Leading Academic Disci-

pline Project of Shanghai Normal University

(DZL706).

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Table1.Templates, preparation and application of material nanochannels

Different templates Membrane preparation applications of special material

of nanochannels nanochannels in analytical

chemistry

membrane template electrochemistry deposition separate the molecular with

emulsion template electroless deposition different size

micromulsion template chemical polymerization separate mixtures containing

other kinds of templates sol-gel chemistry hydrophobic and hydrophilic

chemical vapour deposition molecules

separate the molecular with differ-ent charge

separate proteinDNA


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Figure legends

Figure 1. Cross-section of single -hemolysin channel embedded in a lipid bilayer.4

Figure 2 The transport of goat anti-bovine IgG and goat anti-cat IgG throughAu-Mem and B-IgG-Au-Mem, respectively. 41

Figure 3. Surface SEM images of the membrane after (A) chemical etching and (B) after 5min

(C) 10min (D) 20 min of plasma etching.34


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Figure 1


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Figure 2


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Figure 3


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