표적지향형 나노입자의 농학적 적용

표적지향형 나노입자의 농학적 적용

Agricultural Application of Target Controlled Nanoparticles

Dong-Myung Kim, Ph.D., President

Nanofood Research Society

2008. 9. 26 (Fri)

나노기술 (Nanotechnology)의 역사

1. 1959년 12월 29일 물리학자 Richard Feynman이 캘리포니아 공과대학에서 열린 미 국물리학회 강연에서 처음으로 원자수준의 물질을 다루는 상상을 함.

"There is Plenty of Room at the Bottom"

"Why cannot we write the entire 24 volumes of the Encyclopedia Brittanica on the head of a pin?"

"I am not afraid to consider the final question as to whether, ultimately-in the great future-we can arrange the atoms the way we want; the very atoms, all the way down!"

2. 1974년 노리오 타니구치 (Norio Taniguchi, 도교대학) 교수; 기존의 마이크론 크기의 기술에 대비하여 마이크론 이하(sub-micron)의 크기를 가지는 기술을 “나노기술

(nanotechnology)”

3. 1986년 미국 MIT의 연구원 에릭 드랙슬러 (Eric Drexler)가

"Engines of Creation"이

4. 1998년 한국의 김동명 박사 (서울대학교 나노식품연구회 )가 세계 최초로 식품분야에 나노기술을 도입하고 100nm이하의 크기에서 영양물질을 전달하는 새로운 개념의 식

품을

“나노식품(Nanofood)”이라고 처음 명명하면서 미래의 식량개발의 필요성을 감

조하였다.

Start with a centimeter.

Now divide it into 10 equal parts.

Now divide that into 10 equal parts.

Now divide that into 100 equal parts.

Now divide that into 10 equal parts.

Finally divide that into 100 equal parts.

A centimeter is about the size of a bean.

Each part is a millimeter long. About the size of a flea.

Each part is 100 micrometers long.

About the size (width) of a human hair.

Each part is a micrometer long. About the size of a bacterium.

Each part is a 100 nanometers long.

About the size of a virus.

Each part is a nanometer. About the size of a few atoms or a small molecule.

How small is a nanometer?

1 cm

1 mm

100 μm

1 μm

100 nm

1 nm

Why Nano ???

In bio field, especially, They are similar in size to large biological molecules ("biomolecules") such as Enzymes and Receptor.

- Faster

- Lighter

- Can get into small spaces

- Cheaper

- More energy efficient

- Different properties at very small scale

1. Photo stability 1. Photo stability

General Organic Dye General Organic Dye

Time Time

Intensity Intensity

Nanoparticles For Target Cell

‘S. Nie et al. Nature Biotech., 2004, 22, 969.’

선택성을 가지는 나노 입자

Ideal Concept of Nanoparticles for Medicine

Triple ”F” properties

”find, fight and follow” concept

Highly specific cell targeting molecule Molecule rendering the

particle inert to RES clearance

Contrast agent for monitoring of therapeutic

effect (follow)

Contrast agent for imaging on the level of a single cell (find)

Therapeutic drug (fight)

Molecule promoting endocytosis in target

cell Intelligent biomaterial

Alexa Fluor® 488 FluoroNanogold:

(left) Structures Alexa Fluor® 488 FluoroNanogold-Fab' and streptavidin conjugates, showing separate covalent attachment of the two labels. (right) Fluorescent staining obtained using combined combined Alexa Fluor®* 488 and Nanogold® - Fab' tertiary probe.

Takizawa, T., and Robinson, J. M.:

Use of 1.4-nm immunogold particles for immunocytochemistry on ultra-thin cryosections. J. Histochem. Cytochem., 42, 1615-1623 (1994).

표적지향형 gold nanoparticle

Figure 1: Structure of Palmitoyl NANOGOLD®

Patrick Barger et al. J. Chem. Edu. 1999, 76, 943.

a stable colloidal suspension of sub-domain magnetic particles in a liquid carrier

The particles, which have an average size of about 100Å (10 nm), are coated with a stabilizing dispersing agent

(surfactant) which prevents particle agglomeration even when a strong magnetic field gradient is applied to the ferrofluid.

The surfactant must be matched to the carrier type and must overcome the attractive van der Waals and magnetic forces between the particles. The colloid and thermal stabilities, crucial to many applications, are greatly influenced by the choice of the surfactant. A typical ferrofluid may contain by volume 5% magnetic solid, 10% surfactant and 85% carrier.

Ferrofluid

Super para-magnetic field 10 nm

Fluorescent Magnetic Nanoparticles

Advantages !!!!

-. Dual Function (Magnetic and Fluorescent) -. Core-Shell type (Silicon) -. Bio compatibles (Non toxic) -. Mobilization (Gene, Abs) -. Various surface modification -. Fluorescent (red-580, green-480/500 -. Mass Production blue-400/650)

-. Able in vivo application (MRI) -. Water soluble

Synthesis of Fluorescent Magnetic Nanoparticles (FMN)

Co ferrite magnetic nanoparticle

MNP-SiO₂ Core-shell nanoparticle

TEOS

Incorporation of Organic dye Into the silica shell

MNP@SiO₂(RITC) MNP@SiO₂(FITC) RITC

O N⁺ Cl^- N

COOH CNH

S NH

Si EtO EtO OEt

FITC

O O

NH OH

C NH

S Si

OEt OEtOEt

Ab conjugation with FMN for Separation and Targeting

Schematic illustration of the steps involved in antibody modified MNP@SiO2(FITC) synthesis.

a: CoFe2O4 magnetic nanoparticle,

b: MNP@SiO2(FITC) core-shell nanoparticle,

c: MNP@SiO2(FITC)-PEG/NH2 dual-fabricated core-shell nanoparticle, d: Maleimide terminated smart silica core-shell magnetic nanoparticle, e: MNP@SiO2(FITC)-Ab multifunctional nanoparticle for specific binding.

Carboxyl chains Disulfides

heavy chains

Optical microscope images of the targeted SP2/0 floating cells (white dots) moving due to the application of an external magnetic field.

Before (a) and after (b) the application of a magnetic field by external magnet (~ 0.3 T);

the red dotted circle indicates location of the magnet.

The floating cells move fast in the direction of the magnet (supporting information).

The green back ground color is not due to fluorescence

but due to the back light from the optical microscope equipment.

Agricultural Applications

Approaching for Nutrition Physiology

Application for Pathology

Ideal Concept of Nanoparticles for Plants

1. 병 발생 예찰을 위한 정밀시스템 개발 2. 종합방제 체계구축

3. 고부가가치 생물적방제재 개발 4. 친환경 방제효과 검증

5. 새로운 문제 병해 관리 기술 개발

6. 병원균-기주간 상호반응에 의한 유전체 및 기능분석

7. 식물병리 연구의 기초기반 기술 확립 Approaching for

Nutrition Physiology

Application for Pathology

1. 친환경농업기반기술 :

생산성과 환경이 조화되는 토양관리 기술개발 2. 작물영양진단 신기술 개발 및 피해 경감 기술 :

작물의 양분 및 재해생리 대사 연구

작물 환경스트레스 반응 영양진단 기술개발 및 판단지표 설정 작물의 과잉양분 흡수 피해 해석 연구

영양생리장애 발생 원인 구명 및 장애발현 양분흡수 연구 3. 신속 정확한 작물의 영양진단 기술 개발 :

작물의 양분흡수 및 생리대사 기작 구명 작물의 기후환경 변화 반응 영양생리 구명 작물체 세포내 과잉양분 흡수 전류 기작 구명 Fe및 Mn 등 흡수특성과 영양장애 관련 구명

4. 장애발현지 토양조성이 작물의 영양 흡수에 미치는 영향 : 작물의 영양 결핍에 의한 Chlorosis 와Necrosis 발현특성 영농현장의 다요인 분석에 의한 영양생리장애 발생원인 시설재배지 작물의 내염성 탐색 기술개발

염류과잉이 작물체내 Nitrate 및 Sucrose 대사에 미치는 영향 과잉양분의 작물 세포내 흡수전류 기작 구명

Research Applications

Target Controlled !!!!

-. Magnetic and Fluorescent -. Core-Shell type

-. Bio compatibles & Non toxicity

-. Mobilization (positive charge Æ Gene, LigandÆ Abs etc) -. Various surface modification

-. Visible and water soluble

Strategies !!!!

-. Optimum Condition

-. Optimum Concentration, Time, Sample types

(Cell, Tissue, Whole Plant) and Cell Conditions

-. Applications (endocytosis, ATP activation, Proton pump, osmosis, end blocking)

-. Research & Development

9Incorporation of the fluorescent dyes into the nanoparticles enabled to monitor the movement of cells doped with magnetic nanoparticles under an external magnetic field by optical microscopy, suggesting that multifunctional magnetic nanoparticle could be easily realized on the in vivo target-of-interest and could be ultimately applied to non-invasive delivery systems such as those used in in vivo gene or drug delivery.

9 Nanoparticles doesn’t reveal their toxicity in vitro and in vivo test. So, it can apply to biotechnology have more safety.

9 Cells that uptake or binding nanoparticles are can move by magnetic force.

It prove nanoparticles concentration to specific location by magnet.

9 Positive charged nanoparticles are possible to apply as gene or drug delivery carrier. And it can overcome carrier on a commercial scale limitation in safety, and efficiency.

9As specific ligands and antibody apply to nanoparticles. It is conferred specificity.

In agricultural application, this property will help the research of homing and sorting of plant cells, tissues, and organs.

Conclusions