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## Acción
### URL
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## Hotspot
### Texto
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## Media
### Imágen
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### Video
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## Skin
### Botón
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### Etiqueta
Label_0DD14F09_1744_0507_41AA_D8475423214A.text = LOREM IPSUM
Label_0DD1AF09_1744_0507_41B4_9F5A60B503B2.text = dolor sit amet, consectetur
### Imágen
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### Texto Multilínea
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Peptides, solid- phase synthesis and
characterization: tailor-made methodologies.
Contact
fanny.guzman@pucv.cl
constanza.cardenas@pucv.cl
___
location
address line 1
address line 2
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___
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Integer gravida dui quis euismod placerat. Maecenas quis accumsan ipsum. Aliquam gravida velit at dolor mollis, quis luctus mauris vulputate. Proin condimentum id nunc sed sollicitudin.
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___
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dolor sit amet
consectetur adipiscing elit. Morbi bibendum pharetra lorem, accumsan san nulla.
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___
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dolor sit amet
consectetur adipiscing elit. Morbi bibendum pharetra lorem, accumsan san nulla.
Mauris aliquet neque quis libero consequat vestibulum. Donec lacinia consequat dolor viverra sagittis. Praesent consequat porttitor risus, eu condimentum nunc. Proin et velit ac sapien luctus efficitur egestas ac augue. Nunc dictum, augue eget eleifend interdum, quam libero imperdiet lectus, vel scelerisque turpis lectus vel ligula. Duis a porta sem. Maecenas sollicitudin nunc id risus fringilla, a pharetra orci iaculis. Aliquam turpis ligula, tincidunt sit amet consequat ac, imperdiet non dolor.
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• Sollicitudin tempor sit amet non urna.
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Cleavage
Once the synthesis is finished peptide must be cleaved from the resin.
Step 1. Place each tea bag in a tube correctly labeled.
Step 2. Add cleavage solution, 10 ml per gram of resin.
Step 3. Close the tubes hermetically and incubate, shaking for 3 hours.
Step 4. Precipitate the peptide with cold ether and centrifuge.
Step 5. Perform at least 4 more washes with cold ether, centrifuging and discarding the supernatant.
Step 6. Dry and dissolve milliQ in water, freeze and lyophilize until use. Lyophilized peptide.
Deprotection and Washes
In tea bags protocol, the washings and deprotection steps are done in a single container, changing the solvents used for washing: DMF, IPA and DCM. Deprotection step is accomplished using 20% piperidine in DMF and a solution of BPB is used for monitoring the process colorimetrically.
.
Coupling
Step 1. Preparation of amino acid solutions with activator.
Step 2. Organization of tea bags according to the working document. Double checking.
Step 3. Introduce the tea-bags in their respective container and add the corresponding volume of DIPEA.
Step 4. Coupling, shake for 3 hours. The end of coupling is verified by the absence of blue color of the resin.
Reactor Size
Photo 1: The size of the reactors depends on the system scale, being possible to use from milligrams to grams of resin on a laboratory scale.
Photo 2: The three synthesis protocols presented in this work use different reactors. 40 mg Teabags, 140 mg Liberty Blue and 5 g manual reactor.
Manufacture Tea Bags
Photo 1. Template printed on polypropylene mesh fabric for making the tea bags.
Photo 2. sealing and resin filling of tea-bags.
Photo 3. Tea bags ready to enter the synthesis process.
.
Mass spectrometry analysis.
Photo 1.
Equipment: LCMS-2020 ESI-MS (Shimadzu Corp., Kyoto, Japan).
Mass spectrometry was used to confirm the molecular mass of each product obtained.
Photo 2.
Peptide NBC 112.
MW 1520,8 g/mol.
The three protocols make it possible to obtain the desired product.
Photo 3.
Peptide NBC759.
MW 1742,2 g/mol.
The three protocols make it possible to obtain the desired product.
.
Peptide characterization by HPLC.
Photo 1. Equipment: RP-HPLC, JASCO Corp., Tokyo, Japan.
Peptides were characterized by reversed phase-high-performance liquid chromatography by using a gradient of 0%–70% of solvent B (acetonitrile with 0.05% TFA) versus solvent A (water with 0.05% TFA) on a XBridge™ BEH C18 column (100 × 4.6 mm, 3.5 µm) (Water Corp., Milford, MA, USA), at 1 mL/min flowrate for 8 min.The chromatograms show the differences in purity between the protocols performed.
Photo 2. Peptide NBC112.
The chromatograms show the differences in purity between the protocols performed. Purity is calculated according to the area under the main peak curve.
Photo 3. Peptide NBC759.
The chromatograms show the differences in purity between the protocols performed.Purity is calculated according to the area under the main peak curve.
Solid phase synthesis of the
experimental process
Photo 1: Three synthesis protocols follow the same steps.
The synthesis of the manual reactor using 5 g of resin.
Photo 2: The automated synthesis was carried out with a CEM Liberty blue equipment.
Photo 3: Finally, the simultaneous synthesis protocol in tea bags will be shown in detail next.
Electronic
Journal of
Biotechnology.
Electronic Journal of Biotechnology ISSN 017-3458 is an international, scientific, gold open access journal published every two months which covers all areas related to biotechnology. It is indexed in BIOSIS Previews, Directory of Open Access Journals (DOAJ), INSPEC, Science Citation Index Expanded and Scopus.
Production and Hosting by Elsevier B.V. on behalf of Pontificia Universidad Católica de Valparaíso. Peer review under the responsibility of Pontificia Universidad Católica de Valparaíso.
CONTACT: edbiotec@pucv.cl