Ole auala fa'amavae talafeagai o se mea taua mo le manuia o su'ega peptide. O le faʻamavaeina le talafeagai o le a mafua ai le leiloa o peptides ma le toilalo o le suʻega. Peitai, o le sailia o le auala fa'amavaega lelei o nisi taimi e fai si faigata. O lo'o taua i lalo auala fa'ata'apeina na fa'atūina e vaega fa'apitoa e lua o Suiga MOL. auala
MOL Suia Tekinolosi Group Ta'iala Ta'iala
O le faʻafitauli autu i le faʻamavaeina o le peptide o le faʻavaeina o le fausaga lona lua.
E ui o le faʻavaeina o fausaga lona lua o filifili peptide hydrophobic e sili atu ona manino, vagana ai filifili peptide sili ona pupuu, e tupu lenei mea i le toetoe lava o filifili peptide uma, e tusa lava po o le polarity. O lea la, o le mataupu faavae muamua o le faʻamavaeina polypeptides o le faʻaaogaina o vai faʻamaʻi faʻamaʻi poʻo vai Deionized (po'o vai e leai se okesene pe a fa'ataga).
Ole vaifofo ole peptide e ono fa'afefeteina siama. Ina ia aloese mai lenei tulaga, e tatau ona solo le peptide i vai fa'amama fa'ama'i, po'o le fa'amama i se fa'amama fa'amama fa'atasi ai ma le tele pore o 0.45 pe 0.2 μm mo le faʻamaʻaina. Polypeptides e iai Cys, Feiloai, ma Trp e sili ona a'afia i fa'ama'i, o lea e tatau ai ona fa'amuta i vai e leai se okesene. E mafai ona maua le vai e leai se okesene e ala i le fa'amama i lalo o le fa'aitiitia o le mamafa e ala i le tu'iina o kasa fa'amama (nitrogen, helium, ma argon).
Afai e le mafai ona solu le peptide i le vai mama, togafitiga ultrasonic e mafai ona fesoasoani e talepe vaega ma faʻateleina solubility. Manatua: Ole togafitiga ole Ultrasonic e mafua ai le vevela ole fofo ma le faʻaleagaina ole peptide.
Afai o le peptide o loʻo i ai le tele o amino acids, fa'aaoga 1~10% acetic acid vaifofo vai; mo peptides hydrophobic tele, fa'aoga 50% acetic acid.
Afai o le polypeptide o loʻo i ai se aofaiga tele o acid amino acids, e mafai ona soloia i se 1 ia 10% vaifofo ammonia po'o se pa'u alkaline fe'avea'i pei ole ethyl morphine acetate po'o le bicarbonate. E tatau ona fetuuna'i le tau o le pH a'o le'i faia le chromatography.
Afai o le peptide e tatau ona uta i luga o le koluma, e tatau ona la'ititi le aofa'i o so'o so'o, a leai o le a matua a'afia ai le taimi fa'atumauina.
Afai o le polypeptide e maualuga le hydrophobic aua o loʻo i ai ni filifili pito hydrocarbon manogi e pei o Val, Leu, Feiloai, Fai mai Phe, Tyr, Ala, ma isi., po'o se peptide le mautonu, o le faʻaaogaina o denaturants membrane e pei ole DMF poʻo le DMSO e mafai ona fesoasoani e faʻaumatia le polypeptide:
a. O le maualuga o le faʻaogaina o le membrane denaturants e fesoasoani e faʻaumatia le polypeptide e ala i le faʻaleagaina o lona fausaga lona lua.
e. Membrane denaturants e talafeagai mo le saunia o fofo o suʻesuʻega peptide, ae e ono fa'alavelave i su'esu'ega i a latou gaioiga fa'aola.
i. DMF o le denaturant sili ona lelei (le maualuga maualuga e mafai ona oʻo i ai 30%), ma fa'aopoopo fa'atete'e se'ia fa'amavae le peptide.
o. I le taimi o chromatography fa'asolosolo fa'asolosolo, DMF o le a tafe faʻatasi ma le pito i luma o le eluent. Fa'alagolago ile tele ole tui, atonu e maualuga tele le tau maualuga. O le tele o peptide e mafai ona faʻafefe i totonu o ni nai minute pe a uma le tele o le DMF elutes. Afai e laʻititi le filifili peptide ma vave alu ese, o le a faʻaitiitia le aofaʻi o le peptide.
MOL Suiga Vaega Fa'apitoa II Ta'iala Ta'ape
E fa'afefea ona iloa pe fa'aitutū amino acids, acidic pe alkaline, ma fuafua le auala faataapeina:
1. Tofia uma acid amino acid i le faasologa polypeptide e aofia ai aspartic acid Asp (D), glutamic acid Glu (E) ma C-terminal carboxyl-COOH pei -1; amino acid faavae taitasi e aofia ai arginine Arg ( R), lysine Lys (K), histidine Lana (H) ma le N-terminal amino-NH2 o loʻo tuʻuina atu i ai se tau o +1; amino acids e le tutusa 0. Faʻavae i luga o lenei, le tau o le polypeptide atoa ua fuafuaina, ma le togi lelei Peptides ma se togi leaga ua taʻua o peptides faavae, oi latou e i ai se togi leaga e ta'ua o le acidic peptides, ma oi latou e leai se togi e taʻua o peptides le mautonu.
2. Afai o se peptide alkaline, try to dissolve it with water first; if it cannot be dissolved, try to use 10% – 30% acetic acid; if the peptide is still insoluble, try to use pure acetic acid and trifluoroacetic acid TFA (<50 μl) to dissolve it, and then Dilute the peptide solution to the desired concentration.
3. If it is an acidic peptide, try to dissolve it with water first; if it cannot be dissolved with water, try to use 13% ammonia water (v/v) to dissolve it, and dilute the peptide solution to the required concentration. If the peptide sequence contains cysteine Cys (C), it cannot be dissolved in alkaline solution. Please try the methods below.
4. Neutral peptides are usually dissolved in organic solvents. Tulaga tasi, try to use acetonitrile (acetonitrile), or methanol (methanol) or isopropanol (isopropanol) to dissolve; mo peptides hydrophobic tele, first dissolve with a small amount of dimethyl sulfoxide (DMSO) and dilute with water to Required concentration; if the peptide sequence contains cysteine Cys (C), use dimethylformamide (DMF) poʻo le N-methylpyrrolidone (NMP) mo le faataapeina. Mo peptides e faigofie ona faʻapipiʻi, fa'aopoopo muamua le 6M guanidine·HCl po'o le 8M urea (Urea) e faataape ai, ona fa'asusu lea i le fa'atonuga mana'omia.
fa'aali:
1. Ina ia faafaigofie le teuina ma le faaaogaina mulimuli ane, e fautuaina e faʻamavae le peptide i se faʻatonuga e uiga i 1-2 mg/ml.
2. Ole pa'u fa'amago fa'aisa peptide masani e mafai ona teuina ile -20°C mo le sili atu ma le tasi le tausaga. Mo peptides faʻamavae, e fautuaina e aliquot ma teu i le -20°C; pe afai o le peptide sequence o loʻo i ai le methionine Met (M), Cysine Acid Cys (C) po o le tyrosine Tyr (Y) e fautuaina e teu i totonu o se siosiomaga e leai se okesene e puipuia ai le faʻamaʻiina.
Lisi amino acid fa'apipi'i:
1.Ole amino acids: glycine, alanina, leucine, isoleucine, valine, cystine, methionine, threonine, serine, phenylalanine, tyrosine, tryptophan acid, proline, asparagine ma le glutamine
Fetures: O lenei ituaiga mole mole amino acid e na'o le tasi le vaega amino ma le tasi vaega carboxyl
2.acid amino acids: glutamic acid, aspartic acid
Vaega: O lenei ituaiga mole mole amino acid e aofia ai le tasi vaega amino ma lua vaega carboxyl
3.Amino acids faavae: lisine, arginine, histidine
Vaega: O lenei ituaiga amino acid o loʻo i ai se vaega diaminocarboxyl i lona mole; o le histidine e iai le mama nitrogen ma e vaivai le alkaline, ma o se amino acid faavae foi.
