친수성 코팅 글 리아 딘 - 시아 노 아크릴 레이트 나노 입자의 생산을위한 프로토콜
Summary
This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic. The produced nanoparticle is an assembly of gliadin-cyanoacrylate diblock copolymers. Spray coating with the produced nanoparticle changes the surface of target material to a hydrophilic surface.
Abstract
이 문서는 간단한 스프레이 코팅함으로써 친수성으로 소수성 표면을 변경 단백질 계 나노 입자의 생산을위한 프로토콜을 제공한다. 이러한 나노 입자는 곡류 단백질 (글 리아 딘) 분자의 표면 상에 알킬, 시아 노 아크릴 레이트의 중합 반응에 의해 제조된다. 시아 노 아크릴 레이트는 알킬기가 물질의 표면에인가 될 때 즉각적으로 RT에서 중합 단량체이다. 그 중합 반응은 수분을 포함하는 표면에 약 염기성 또는 구핵 종의 미량으로 개시된다. 중합 후에 니트릴 기는 폴리 (알킬 시아 노 아크릴 레이트)의 중추이기 때문에, 중합 된 알킬 시아 노 아크릴 레이트는 오브젝트 물질과 강한 친 화성을 나타낸다. 그들은 시아 노 아크릴 레이트의 중합을 개시 할 수 아민 그룹을 포함하기 때문에 단백질이 중합 개시제로서 작동합니다. 응집 된 단백질은 개시제로서 사용되는 경우, 단백질 응집물은 소수성 둘러싸여폴리 (알킬 시아 노 아크릴 레이트) 알킬, 시아 노 아크릴 레이트의 중합 반응 후의 체인. 실험 조건을 제어함으로써, 나노 미터 범위의 입자가 제조된다. 생성 된 나노 입자를 용이하게 유리, 금속, 플라스틱, 목재, 피혁, 직물을 포함하여 대부분의 재료의 표면에 흡착. 재료의 표면이 생성 된 나노 입자 현탁액을 분무하고, 물로 세정하는 경우, 나노 입자의 미셀 구조의 형태를 변경하고, 친수성 단백질은 공기에 노출된다. 그 결과, 나노 입자 표면에 코팅 된 친수성으로 변화시킨다.
Introduction
The goal of this article is to show the protocol for the preparation of nanoparticle suspension that modifies the wetting property of materials by a simple spray. The presented nanoparticle suspension is made from alkyl cyanoacrylate1 and a cereal protein, gliadin2,3. During the manufacturing process, protein aggregates are formed in aqueous ethanol4. Subsequent reaction with monomer (alkyl cyanoacrylate) produces the nanoparticle that is comprised of a protein core surrounded by linear polymer chains [poly(alkyl cyanoacrylate)]5.
Poly(alkyl cyanoacrylate)s are biodegradable and have been used for the production of nanoparticles via emulsion polymerization6. This reaction is spontaneously initiated by the hydroxyl groups dissociated from water or by other nucleophilic groups in the reaction medium7. In the case of the reaction presented in this article, the amine groups on the surface of protein aggregates initiate the polymerization reaction of alkyl cyanoacrylate monomers5,8. As a result of this reaction, nanoparticles are formed in the reaction medium. The core of the nanoparticle is protein aggregates and the outer layer is poly(alkyl cyanoacrylate) (PACA) chains. The prepared nanoparticle has a strong affinity on most materials (more precisely, any material which PACA can adsorb to) and adheres onto their surface to form a thin coating on a nanometer scale. A simple spray coating instantly turns the surface of the materials hydrophilic.
Gliadin is one of the main fractions of gluten, which is in the endosperms of wheat. Gliadins are mainly monomeric proteins with molecular weights around 28,000 – 55,000. Non-covalent bonds such as hydrogen bonds, ionic bonds and hydrophobic bonds are responsible for the aggregation of gliadins2. Although gliadin is chosen as a reactant in this article, many other proteins can also be used for the same purpose. However, the reaction condition needs to be modified accordingly because the condition for inducing aggregation is dependent on the type of protein to be employed8. Compared with other proteins, gliadin is more readily available, purification is simple, and production cost is low. Although ethyl cyanoacrylate (ECA) is chosen as a monomer for the presented reaction, other alkyl cyanoacrylates can also be used for the same reaction. The reason for choosing ECA is that it is readily available at low cost.
Protocol
Representative Results
Discussion
There are several critical steps in the production of the nanoparticle suspension. If the purified gliadin contains impurities, the reaction with ECA will produce side products. Although these unwanted products can be removed from the reaction medium during the centrifugation stage, it lowers the yield of the major product. If the gliadin solution prepared during experimental step 2.3) does not show clear separation between supernatant and precipitate after two days, the solution needs to stand for longer time. Using fre…
Offenlegungen
The authors have nothing to disclose.
Acknowledgements
전문가 기술 지원 씨 제이슨 아드 킨스에게 감사드립니다.
Materials
| Ethyl cyanoacrylate (ECA) monomer | K&R International (Laguna Niguel, CA) | I-1605 | Any pure ECA can be used. |
| Gliadin | MGP Ingredients, Inc (Atchison, KS) | Gift from the company | Gliadin can be purchased from Sigma-Aldrich (cat #: G3375-25G). Instead of gliadin, any commercial gluten can be used. |
| HCl | Any | Any reagent grade chemical can be used. | |
| Acetone | Any | Any reagent grade chemical can be used. | |
| Methanol | Any | Any reagent grade chemical can be used. | |
| Ethanol (100%) | Any | Any reagent grade chemical can be used. | |
| Filter paper | Any | Any grade filter paper larger than 10 cm can be used. | |
| Cell culture square dish | Any | Any dish larger than 20 cm x 20 cm can be used. | |
| Coffee grinder | Any | Any coffee grinder can be used. | |
| Rotary evaporator | Any | Any rotary evaporator can be used. | |
| Freeze Dryer | Any | Any freeze dryer that can reach – 70°C can be used. | |
| Centrifuge | Any | Any centrifuge that can apply 1000 x g can be used. | |
| Magnetic stirrer | Any | Any magnetic stirrer that can turn spin bar to 1000 RPM can be used. | |
| Dynamic Light Scattering (DLS) | Brookhaven Instruments Corporation | NanoBrook Omni Zeta Potential Analyzer | DLS from any company can be used. |
| Scanning Electron Microscope (SEM) | Carl Zeiss Inc. | Any SEM can be used. | |
| Dynamic Contact Angle (DCA) | Thermo Cahn Instruments | Any DCA can be used. |
Referenzen
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