Nyuaj Peptide Synthesis|Hydrophobic Peptide Customization|Nyuaj Peptide Synthesis|Cluster Sequence Synthesis|Kev tshawb fawb-Peptide
Meta Description
Peptide sequence dhau hydrophobic? Aggregate yooj yim? Ntau khub ntawm disulfide bonds? Kev tshawb fawb-Peptide's Difficult Peptide Synthesis thiab Hydrophobic Peptide Synthesis cov kev pabcuam tshwj xeeb hauv txhua yam "tsis tuaj yeem ua tiav". 20 xyoo dhau los, cov tshuab hnyav tshwj xeeb, xa khoom ruaj khov. Thov koj xav tiv tauj peb.
Qee cov peptides yog qhov nyuaj
Dab tsi ntawm peptides raug suav hais tias yog "nyuaj"?
Tau nyob hauv kev lag luam peptide synthesis rau 20 xyoo, kuv tau pom txhua yam nyuaj peptides. Thaum cov neeg siv khoom tuaj rau peb, lawv tau nug ntau lub tuam txhab, thiab cov lus teb lawv tau txais yog "Kuv ua tsis tau" lossis "sim nws, tsis muaj kev lees paub".
Cov peptides twg tau yooj yim tsis lees paub?
Peptide nrog muaj zog hydrophobicity: transmembrane cheeb tsam, teeb liab peptide, hydrophobic amino acid, thaum synthesized, nws yuav clumped ua ke ntawm resin, thiab cov amino acid qab nws yuav tsis tau nkag mus.
Peptides uas yooj yim aggregated, xws li amyloid thiab antimicrobial peptides, yog piled thaum synthesized thiab clog lub kem thaum purified.
Peptides nrog ntau khub ntawm disulfide bonds: ntau tshaj peb khub, ib kauj ruam tsis ncaj ncees lawm nyob rau hauv kev txiav txim khub thiab tag nrho cov molecule puas.
Ntev peptide: ntau dua 60 amino acids, txhua kauj ruam ntawm cov txiaj ntsig tau txo qis, thiab thaum kawg tsis muaj dab tsi sab laug.
Kev sib xyaw ua ke ntawm kev hloov kho: phosphorylation, fluorescence, cyclization, thiab tej yam kev mob uas sib ntaus sib tua.
Peb tuaj yeem coj lawv txhua tus, thiab nws tsis yog "sim", nws yog "muaj phiaj xwm".
Peb yuav ua li cas
1. Hydrophobic peptides: nrog tshwj xeeb hnyav tshuab
Qhov nyuaj ntawm hydrophobic peptides nyob rau hauv qhov tseeb tias lawv tsis tuaj yeem yaj hauv cov kuab tshuaj, thiab thaum ua ke, cov saw peptide "clumps ua ke" ntawm cov resin, ua rau nws tsis yooj yim rau cov amino acids tom qab nkag mus.
Peb qhov kev daws teeb meem:
Kev sib xyaw ua ke tshwj xeeb: DMF, NMP, DCM sib xyaw ua ke, ntxiv rau me ntsis kua tso tawm tus neeg sawv cev, kom cov saw peptide tau khaws cia ntev.
Kev ntsuas kub kom zoo: qee qhov kev cuam tshuam zoo dua ntawm 40℃dua li chav tsev kub, qee qhov yuav tsum tau qis dua 0 degree, peb tau sim lawv ib qho los ntawm ib qho.
Resin xaiv: Siv cov dej solubility siab rau hydrophobic peptides kom tawm qhov chaw txaus rau peptide chains.
Solubilized labels: Yog tias koj ua tsis tau, ntxiv ib daim ntawv lo hydrophilic thiab txiav nws tom qab kev sib txuas.
Cov ntaub ntawv tiag tiag: ib qho 35aa transmembrane peptide, hydrophobic amino acids accounted rau 70%, tus neeg siv khoom nrhiav peb lub tuam txhab ua tsis tau. Peb hloov cov kuab tshuaj thiab ntxiv solubilizing daim ntawv lo, thaum kawg lub purity yog 92%, thiab cov neeg muas zaub ua tus qauv kawm smoothly.
2. Ntau khub ntawm disulfide bonds: tsis muaj hmoo, zoo
Qhov ntau disulfide bonds muaj, qhov nyuaj nws yog ua ke lawv. Ob khub zoo, tab sis peb lossis ntau dua, thiab ib kauj ruam tsis raug thiab nws tag nrho.
Peb lub tswv yim:
Kev tiv thaiv Orthogonal: Cys tuaj yeem qhib thiab ua khub hauv cov kauj ruam uas siv cov pab pawg tiv thaiv sib txawv, xws li Trt, Acm, tBu.
Optimization ntawm oxidation tej yam kev mob: huab cua oxidation, glutathione oxidation, iodine oxidation, txawv txoj kev sib txawv peptides.
Intermediate QC: Tom qab txhua tus khub, coj tus qauv rau MS thiab paub meej tias kev ua khub ua ntej mus.
MS/MS kev pom zoo (xaiv tau): kev so zaum kawg- nrog rau qhov thib ob pawg spectrometry kom pom qhov twg cov nyiaj disulfide tau txuas.
A real case: A conotoxin peptide containing four pairs of disulfide bonds was rejected by many companies. We designed a four-step oxidation strategy, which took six weeks, and delivered >90% purity, raug muab sib koom ua ke disulfide daim ntawv cog lus, thiab cov qauv siv lead ua tau raug daws los ntawm cov neeg siv khoom.
3. Kev sib sau ua ke -peptides nquag: ua ke thaum tiv thaiv
Qee cov peptides ib txwm nyiam sib sau ua ke, xws li amyloid protein thiab tshuaj tua kab mob peptide. Lawv pib sib sau ua ke thaum lawv sib txuas, thaiv kab ke thaum lawv raug ntxuav, thiab tsis tuaj yeem yaj thaum lawv nyob hauv tes.
Peb qhov kev daws teeb meem:
Pseudoproline: Pseudoproline tau qhia ntawm Ser / Thr txoj haujlwm los cuam tshuam kev sib sau ua ke thiab hloov rov qab tom qab kev sib txuas.
Segmental synthesis: Peptides uas yog polymerized dhau lawm raug txiav mus rau hauv ob peb ntu thiab ua ke sib cais, thiab tom qab ntawd rov txuas dua thaum kawg.
Kev ua kom zoo ntawm kev ua kom huv huv: kub siab, acidic pH, ntxiv cov kuab tshuaj organic kom pom cov xwm txheej uas ua rau nws tsis yog -polymerizing.
4. Complex modifier ob peb ua ke: tej yam kev mob uas tsis muaj fights
Qee cov peptides muaj ntau yam yuav tsum tau ua: phosphorylation, fluorescent labeling, thiab cyclization. Cov kev hloov kho no yuav tsis sib haum xeeb - piv txwv li, phosphorylation yog acid-phobic, fluorescent labeling yog lub teeb-phobic, thiab cyclization yuav tsum muaj pH tshwj xeeb.
Peb txoj hauv kev:
Kev txuag lub tswv yim tsim: uas trim ua ntej, uas trim ua tom qab, tsim ua ntej.
Orthogonal tej yam kev mob: Cov xwm txheej ntawm txhua kauj ruam ntawm cov tshuaj tiv thaiv tsis cuam tshuam rau cov kev hloov kho uas twb tau ua lawm.
Intermediate QC: Tom qab txhua kauj ruam ntawm kev hloov kho, coj tus qauv kom paub tseeb tias nws ua tiav kom raug ua ntej tsiv mus.
5. Purity: Nyob ntawm seb koj ua dab tsi nrog nws
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Purity theem |
Siv tau Scenarios |
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Cov pa ntshiab (85-95%) |
Ua ke nrog kev sim, kev tshuaj ntsuam thawj zaug |
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Siab purity (95-98%) |
cellular thwmsim, functional validation |
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Ultrapure (>98%) |
Kev tshawb fawb txog cov qauv, crystallography |
Txhua peptide yog nrog los ntawm HPLC thiab MS daim ntawv qhia, ua kom cov ntaub ntawv purity pob tshab.
Vim li cas thiaj hu peb rau Nyuaj Peptide Synthesis?
1. Peb tau ua qhov no tau 20 xyoo
Nyuaj Peptide Synthesis tsis yog ib yam uas leej twg ua tau. Tau ntau xyoo, peb tau ua ntau yam nyuaj peptides: transmembrane proteins, viral fusion peptides, conotoxins, amyloid proteins, antimicrobial peptides, thiab lwm yam. Peb muaj ib lub tswv yim zoo ntawm cov sequences muaj teeb meem, thiab tej yam kev mob yuav cawm tau lawv. Tsawg kawg koj tsis tas yuav sim txhua tus ntawm lawv yog tias koj tuaj rau peb rau kev nyuaj thiab Hydrophobic Peptide Synthesis.
2. Nws tsis yog "sim", nws yog "muaj kev pab cuam".
Ntau lub tuam txhab siv cov peptides nyuaj thiab hais tias, "Peb yuav sim nws, tab sis peb tsis lav nws". Peb tsis ua si li ntawd. Ua ntej lees txais ib qho kev txiav txim, peb yuav ntsuas qhov sib lawv liag, qhov kev faib ua feem ntawm hydrophobic amino acids yog dab tsi, seb puas muaj kev paub txog kev sib sau ua ke, muaj pes tsawg khub ntawm disulfide bonds, thiab seb qhov kev hloov kho puas nyuaj lossis tsis. Tom qab kev ntsuam xyuas, peb yuav qhia rau koj: dab tsi yog qhov kev vam meej thiab ntev npaum li cas yog lub sij hawm voj voog.
Peb tsis muab cov xaj uas ua tsis tau, thiab peb yuav tsum xa cov xaj uas peb tau txais.
3. Tshwj xeeb hnyav tshuab rau hydrophobicity
Hydrophobic peptide yog peb lub ntsiab lus muaj zog. Yog tias cov kuab tshuaj tsis tuaj yeem yaj, peb yuav hloov cov kuab tshuaj; yog tias qhov kub thiab txias tsis hnov tsw, peb yuav kho qhov kub thiab txias; yog tias cov pa roj hmab tsis tuav tawm, peb yuav hloov cov resin. Tseem muaj cov ntawv solubilizing raws li txoj kev rov qab tawm.
4. Ntau khub ntawm disulfide bonds, kauj ruam -los ntawm - kauj ruam
Yog tias muaj ntau tshaj peb khub ntawm disulfide bonds, peb yuav ua ib kauj ruam -los ntawm - kauj ruam mus kom ze: thawj kauj ruam yog los tiv thaiv A thiab qhib thawj khub; tiv thaiv B thiab qhib ob khub; tiv thaiv C thiab qhib tus khub thib peb. Tom qab txhua kauj ruam, peb coj cov qauv rau MS thiab paub meej tias kev sib koom ua ke ua ntej yuav mus. Thaum kawg, MS / MS yuav tawg thiab txheeb xyuas, thiab cov ntaub ntawv yuav muab rau koj yam tsis muaj koj twv.
5. Kev tswj kom zoo: cov ntaub ntawv ntawm txhua kauj ruam
Kev tswj kom zoo ntawm peptides nyuaj yuav tsum tau ceev faj ntxiv:
Intermediate QC: Tom qab txhua ntu thiab txhua kauj ruam ntawm kev hloov kho, ntsuas qhov purity thiab molecular hnyav ua ntej.
Cov khoom kawg: HPLC rau purity, MS rau molecular hnyav.
Solubility test: Muab cov kuab tshuaj pom zoo thiab cov txheej txheem solubilization, kom koj tsis tau txais peptides uas tsis yaj.
Ua nyuaj thiab Hydrophobic Peptide Synthesis, cov ntaub ntawv hais lus rau nws tus kheej, txhua kauj ruam tuaj yeem taug qab.
Qhov twg tuaj yeem siv cov Peptides nyuaj?
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Cov chaw tshawb fawb |
Hom nyuaj peptide hom |
Peb tau ua qhov xwm txheej qhov twg |
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membrane proteins |
transmembrane cheeb tsam, hydrophobic nplhaib |
Ntau cheeb tsam transmembrane, hydrophobic amino acids> 60% |
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Cov kab mob neurodegenerative |
A , -synuclein, Tau |
Yooj yim aggregated, nyuaj rau purify |
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kev tshawb nrhiav tshuaj lom |
Conotoxin, nab venom peptides |
Ntau khub ntawm disulfide bonds, siab ua khub yuav tsum |
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kab mob proteins |
fusion peptides, spiny protein fragments |
Ntev ntev thiab hydrophobicity |
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Antimicrobial peptides |
natural antimicrobial peptides |
Yooj yim aggregated, zoo them |
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cell{0}}peptides nkag mus |
Amphiphilic membrane- nkag mus rau peptides |
Hloov hydrophobic thiab hydrophilic, yooj yim precipitated |
Kev xa khoom thiab kev tswj xyuas zoo
- Daim ntawv xa tuaj: lyophilized hmoov, centrifuge raj lossis vials.
- Cov ntaub ntawv muaj xws li: COA (HPLC profile + MS spectrum), qhia txog kev sib txuas (yeem).
- Kev ntsuam xyuas xaiv tau: peptide cov ntsiab lus, endotoxin, noo noo, amino acid muaj pes tsawg leeg, kev ntsuas solubility.
- Customized Ntim: faib thiab kos npe raws li koj xav tau.
- Txhua qhov nyuaj thiab Hydrophobic Peptide Synthesis cov khoom raug tshuaj xyuas kom ntseeg tau tias qhov ua tiav yog qhov tseeb thiab qhov purity yog raws li tus qauv ua ntej nws raug xa tawm.
Peb lub neej tiag -
Case 1: Ib qho kev pab cuam transmembrane peptide ntawm lub tuam txhab kws tshuaj
Cov neeg siv khoom xav tau 45aa transmembrane peptide nrog 65% hydrophobic amino acids rau cov protein-protein kev sib cuam tshuam kev kawm. Peb pom peb lub tuam txhab, ob ntawm lawv tsis kam lees qhov kev txiav txim, thiab ib qho ntawm lawv muaj tsawg dua 70% purity tom qab peb lub hlis. Tom qab kev ntsuam xyuas, peb tau txais ib qho tshwj xeeb hnyav system nrog solubilizing daim ntawv lo thiab xa cov khoom nyob rau hauv rau lub lis piam nrog 94% purity. Cov neeg siv khoom tau txais cov ntaub ntawv sib khi zoo.
Case 2: Conotoxin project ntawm ib lub koom haum tshawb fawb
Cov neeg siv khoom xav tau ib qho conotoxin peptide uas muaj plaub khub ntawm disulfide daim ntawv cog lus rau kev kawm txog cov qauv. Peb tau tsim ib qho plaub - kauj ruam oxidation lub tswv yim, thiab txhua khub ntawm disulfide daim ntawv cog lus tau coj mus kuaj thiab lees paub los ntawm MS. Cov khoom kawg tau xa nrog 91% purity thiab MS / MS kev pom zoo ntawm qhov raug ua khub. Tus neeg siv khoom siv peptide no los daws peb - qauv qauv, thiab peptide tau tshaj tawm hauv qhov teeb meem saum toj kawg nkaus.
Case 3: amyloid program ntawm ib lub tsev kawm ntawv txawv teb chaws
The customer needed a 62aa fragment of alpha-synuclein to study the aggregation mechanism. This peptide starts to aggregate when it is synthesized and always clogs up the column when it is purified. We introduced pseudoproline to destroy the aggregation tendency, changed it back after synthesis, and optimized the purification conditions, and finally got the product with purity >95%, thiab cov neeg siv khoom tau ua ThT thiab TEM smoothly.
Tham txog koj cov kev xav tau siab peptide?
Txawm hais tias koj tau raug tsis lees paub los ntawm lwm tus, lossis koj tuaj yeem ua tsis tau hauv koj lub chaw kuaj mob, koj tuaj yeem tham nrog peb. Yog tias koj tab tom nrhiav Science-Peptide ua nyuaj thiab Hydrophobic Peptide Synthesis, tsawg kawg koj tsis tas yuav sim ib qho los ntawm ib qho.
Kuv xav kom koj qhia kuv:
Peptide sequence (los yog cov ntaub ntawv phiaj xwm)
Koj puas tau muaj lwm tus ua nws ua ntej? Qhov tshwm sim yog dab tsi?
Ntau npaum li cas ( milligrams)
purity xav tau
Thaum twg koj xav tau?
Muab koj qhov kev ntsuam xyuas ua tau thiab qhov kev tsocai hauv 24 teev.