Name: a protocol for the production of gliadin-cyanoacrylate nanoparticles for hydrophilic coating
Uploaded: 2016-07-08T14:00:00
Description: Watch this Scientific Journal Video about A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating at JoVE.com

وبفضل السيد جيسون آدكنز لتقديم المساعدة التقنية للخبراء.

1. Defatting غليادين التجاري <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> قياس 150 مل من الأسيتون مع الاسطوانة وتصب في 250 مل دورق مخروطي. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> مع التحريك مع شريط تدور على محرك مغناطيسي في RT، إضافة 30 غرام من مسحوق غليادين التجار...

<ol>
	<li>Vauthier, C., Dubernet, C., Fattal, E., Pinto-Alphandary, H., Couvreur, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Poly(alkylcyanoacrylates)+as+biodegradable+materials+for+biomedical+applications.">Poly(alkylcyanoacrylates) as biodegradable materials for biomedical applications.</a> Adv. Drug Deliver. Rev. 55, 519-548 (2003).</li><li>Wieser, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemistry+of+gluten+proteins.">Chemistry of gluten proteins.</a> Food Microbiol. 24, 115-119 (2007).</li><li>Bietz, J. A., Wall, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identity+of+high+molecular+weight+gliadin+and+ethanol+soluble+glutenin+subunits+of+wheat:+Relation+to+gluten+structure.">Identity of high molecular weight gliadin and ethanol soluble glutenin subunits of wheat: Relation to gluten structure.</a> Cereal Chem. 57, 415-421 (1980).</li><li>Kim, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+of+composites+by+using+gliadin+as+a+bonding+material.">Production of composites by using gliadin as a bonding material.</a> J. Cereal Sci. 54, 168-172 (2011).</li><li>Kim, S., Kim, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+of+gliadin-poly(ethyl+cyanoacrylate)+nanoparticles+for+hydrophilic+coating.">Production of gliadin-poly(ethyl cyanoacrylate) nanoparticles for hydrophilic coating.</a> J. Nanopart. Res. 16, 1-10 (2014).</li><li>Behan, N., Birkinshaw, C., Clarke, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Poly+n-butyl+cyanoacrylate+nanoparticles:+a+mechanistic+study+of+polymerisation+and+particle+formation.">Poly n-butyl cyanoacrylate nanoparticles: a mechanistic study of polymerisation and particle formation.</a> Biomaterials. 22, 1335-1344 (2001).</li><li>Nicolas, J., Couvreur, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+poly(alkyl+cyanoacrylate)-based+colloidal+nanomedicines.">Synthesis of poly(alkyl cyanoacrylate)-based colloidal nanomedicines.</a> Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol. 1, 111-127 (2009).</li><li>Kim, S., Evans, K., Biswas, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+of+BSA-poly(ethyl+cyanoacrylate)+nanoparticles+as+a+coating+material+that+improves+wetting+property.">Production of BSA-poly(ethyl cyanoacrylate) nanoparticles as a coating material that improves wetting property.</a> Colloid Surface. B. 107, 68-75 (2013).</li><li>Lander, L. M., Siewierski, L. M., Brittain, W. J., Vogler, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+comparison+of+contact+angle+methods.">A systematic comparison of contact angle methods.</a> Langmuir. 9, 2237-2239 (1993).</li><li>Davies, J., Nunnerley, C. S., Brisley, A. C., Edwards, J. C., Finlayson, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+Dynamic+Contact+Angle+Profile+Analysis+in+Studying+the+Kinetics+of+Protein+Removal+from+Steel+Glass,+Polytetrafluoroethylene,+Polypropylene,+Ethylenepropylene+Rubber,+and+Silicone+Surfaces.">Use of Dynamic Contact Angle Profile Analysis in Studying the Kinetics of Protein Removal from Steel Glass, Polytetrafluoroethylene, Polypropylene, Ethylenepropylene Rubber, and Silicone Surfaces.</a> J. Colloid Interf. Sci. 182, 437-443 (1996).</li><li>Giolando, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nano-crystals+of+titanium+dioxide+in+aluminum+oxide:+A+transparent+self-cleaning+coating+applicable+to+solar+energy.">Nano-crystals of titanium dioxide in aluminum oxide: A transparent self-cleaning coating applicable to solar energy.</a> Sol. Energy. 97, 195-199 (2013).</li></ol>

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...

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.

<table border="0" cellpadding="0" cellspacing="0" width="580">
	<tbody>
		<tr height="85">
			<td height="85" style="height:85px;width:143px;">Ethyl cyanoacrylate (ECA) monomer</td>
			<td style="width:113px;">K&#38;R International (Laguna Niguel, CA)</td>
			<td style="width:107px;">I-1605</td>
			<td style="width:219px;">Any pure ECA can be used.</td>
		</tr>
		<tr height="106">
			<td height="106" style="height:106px;width:143px;">Gliadin</td>
			<td style="width:113px;">MGP Ingredients, Inc (Atchison, KS)</td>
			<td style="width:107px;">Gift from the company</td>
			<td style="width:219px;">Gliadin can be purchased from Sigma-Aldrich (cat #: G3375-25G). Instead of gliadin, any commercial&#160; gluten can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">HCl</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any reagent grade chemical can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Acetone</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any reagent grade chemical can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Methanol</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any reagent grade chemical can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Ethanol (100%)</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any reagent grade chemical can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Filter paper</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any grade filter paper larger than 10 cm can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Cell culture square dish</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any dish larger than 20 cm x 20 cm can be used.</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:143px;">Coffee grinder</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any coffee grinder can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Rotary evaporator</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any rotary evaporator can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Freeze Dryer</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any freeze dryer that can reach - 70&#176;C can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Centrifuge</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any centrifuge that can apply 1000 x g can be used.</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:143px;">Magnetic stirrer</td>
			<td style="width:113px;">Any</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any magnetic stirrer that can turn spin bar to 1000 RPM can be used.</td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:143px;">Dynamic Light Scattering (DLS)</td>
			<td style="width:113px;">Brookhaven Instruments Corporation</td>
			<td style="width:107px;">NanoBrook Omni Zeta Potential Analyzer</td>
			<td style="width:219px;">DLS from any company can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Scanning Electron Microscope (SEM)</td>
			<td style="width:113px;">Carl Zeiss Inc.</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any SEM can be used.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:143px;">Dynamic Contact Angle (DCA)</td>
			<td style="width:113px;">Thermo Cahn Instruments</td>
			<td style="width:107px;"></td>
			<td style="width:219px;">Any DCA can be used.</td>
		</tr>
	</tbody>
</table>

a protocol for the production of gliadin-cyanoacrylate nanoparticles for hydrophilic coating

تقدم هذه المقالة بروتوكول لإنتاج الجسيمات النانوية البروتين على أساس أن يغير سطح مسعور لمحبة للماء عن طريق رذاذ الطلاء بسيط. وتنتج هذه الجسيمات النانوية عن طريق تفاعل البلمرة من cyanoacrylate ألكيل على سطح بروتين الحبوب (غليادين) الجزيئات. الكيل cyanoacrylate هو مونومر أن يبلمر على الفور في RT عندما يتم تطبيقه على السطح من المواد. يبدأ في رد فعل البلمرة من كميات ضئيلة من الأنواع ضعيف الأساسية أو أليفة النواة على السطح، بما في ذلك الرطوبة. مرة واحدة بلمرة، وتظهر cyanoacrylates ألكيل بلمرة صلة قوية مع المواد الكائن لأن الجماعات النتريل هي في العمود الفقري للبولي (ألكيل cyanoacrylate). تعمل البروتينات أيضا المبادر لهذا البلمرة لأنها تحتوي على مجموعة أمين يمكن أن الشروع في بلمرة cyanoacrylate. إذا تم استخدام البروتين تجميع باعتباره المبادر، وتحيط مجموع البروتين من قبل مسعوربولي (ألكيل cyanoacrylate) سلاسل بعد رد فعل البلمرة من cyanoacrylate ألكيل. من خلال السيطرة على حالة تجريبية، يتم إنتاج جسيمات في نطاق نانومتر. النانوية المنتجة كثف بسهولة على سطح معظم المواد بما في ذلك الزجاج والمعادن والبلاستيك والخشب والجلود، والأقمشة. عندما يتم رش سطح مادة مع تعليق جسيمات متناهية الصغر المنتجة وتشطف بالماء، وهيكل micellar من جسيمات متناهية الصغر يتغير التشكل، ويتعرض البروتينات ماء في الهواء. ونتيجة لذلك، يتغير سطح المغلفة جسيمات متناهية الصغر إلى محبة للماء.

النانوية يمكن أن تكون مستعدة في ظروف التفاعل المختلفة. أشكال غليادين تتجمع في مجموعة واسعة من محتوى الإيثانول 5. ومع ذلك، فإن حجم الركام يحتاج إلى أن تكون صغيرة قدر الإمكان لأن طبقة إضافية (أي، بلمرة ECA) ستضاف إلى هذا الركام وهذه العملية تج?...

Watch this Scientific Journal Video about A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating at JoVE.com

A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating

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.

بروتوكول لإنتاج الجسيمات النانوية غليادين-cyanoacrylate لمع الماء طلاء

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic by a simple spray ...

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