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Porous silica particles anakonzedwa ndi Sol-gel osakaniza njira ndi zosintha zina kupeza macroporous particles.These particles anali derivatized ndi reversible Kuwonjezera kugawikana unyolo kutengerapo (RAFT) polymerization ndi N-phenylmaleimide-methylvinylisocyanate (PMI) ndi styrene kukonzekera N-phenylmaleide zitsulo polyborena polyborimide polyboride polyboride polyboride s (100 × 1.8 mm id) anali odzaza ndi slurry kulongedza.Anayesedwa ndime ya PMP kulekanitsidwa kwa peptide osakaniza opangidwa ndi peptides asanu (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arnkeg, leucineps ntchito album) digesinch humano digestixips dicineps album digicineps. .Pansi pazikhalidwe zabwino kwambiri, chiwerengero cha mbale zowerengera za osakaniza a peptide ndipamwamba kwambiri mpaka 280,000 mbale / m². Poyerekeza ntchito yolekanitsa ya gawo lomwe linapangidwa ndi malonda a Ascentis Express RP-Amide, adawona kuti kulekanitsa kwa gawo la PMP kunali kopambana kuposa malonda a malonda ndi kuthetsa kusiyana.
M'zaka zaposachedwa, msika wa biopharmaceutical wakhala msika womwe ukukula padziko lonse lapansi ndikuwonjezeka kwakukulu kwa msika. madzimadzi a m'thupi, minofu ndi maselo ndi ntchito yovuta kwambiri chifukwa cha kuchuluka kwa mitundu yomwe ingathe kuzindikirika mu chitsanzo chimodzi.Ngakhale kuti misa spectrometry ndi chida chothandiza kwa peptide ndi mapuloteni sequencing, ngati zitsanzo zoterezi jekeseni mu spectrometer misa mu chiphaso chimodzi, kulekana sikudzakhala ideal. spectrometer pa nthawi yapatsidwa4,5,6.Kuonjezera apo, panthawi yopatukana yamadzimadzi, owunikira amatha kuyang'ana m'madera opapatiza, potero amaika ma analyticswa ndikuwongolera kuzindikira kwa MS sensitivity.Liquid chromatography (LC) yapita patsogolo kwambiri m'zaka khumi zapitazi ndipo yakhala njira yodziwika bwino mu proteomic analysis7,8,9,10.
Reversed-phase liquid chromatography (RP-LC) amagwiritsidwa ntchito kwambiri kuyeretsa ndi kulekanitsa zosakaniza za peptide pogwiritsa ntchito octadecyl-modified silica (ODS) monga gawo lokhazikika11,12,13. chofunika kuti tifufuze ma peptides ndi mapuloteni okhala ndi polar ndi osakhala polar moieties kuti agwirizane ndi kusunga ma analytes16.Mixed-mode chromatography, yomwe imapereka ma multimodal interactions, ikhoza kukhala njira ina ya RP-LC yolekanitsa ma peptides, mapuloteni, ndi zosakaniza zina zovuta. 8,19,20,21.Mixed-mode stationary phases (WAX/RPLC, HILIC/RPLC, polar intercalation/RPLC) ndi yoyenera kupatukana kwa peptide ndi mapuloteni chifukwa cha kukhalapo kwa magulu a polar ndi omwe si a polar22,23,24,25,26,27,28 kusankha kwa ma analyte a polar ndi omwe si a polar, popeza kulekana kumadalira kuyanjana pakati pa analyte ndi gawo loyima.Kuyanjana kwa Multimodal 29, 30, 31, 32.Posachedwapa, Zhang et al.30 anakonza dodecyl-zitha polyamine stationary gawo ndi bwinobwino analekanitsa hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, ndi angapo analytes.The polar intercalator ali onse polar ndi sanali polar magulu, kotero angagwiritsidwe ntchito kulekanitsa peptides ndi mapuloteni kuti onse hydrophobic ndi Czamos hydrophilicembed hydrophilicembed hydrophilicembed hydrophilamide-molumo. columns) amapezeka pamalonda pansi pa dzina la malonda la Ascentis Express RP-Amide, koma mizatiyi imagwiritsidwa ntchito pofufuza amine 33 kokha.
Pakafukufuku wamakono, polar-embedded stationary phase (N-phenylmaleimide-embedded polystyrene) inakonzedwa ndikuwunikiridwa kuti ilekanitse ma peptides ndi trypsin digests a HSA. Gawo loyima linakonzedwa pogwiritsa ntchito njira zotsatirazi.Tinthu tating'onoting'ono ta silika tinakonzedwa molingana ndi ndondomeko yomwe inaperekedwa m'buku lathu lapitalo ndi zosinthidwa zina, T. cetic acid inasinthidwa kuti ikonzekere tinthu ta silika ndi kukula kwakukulu kwa pore.Chachiwiri, ligand yatsopano, phenylmaleimide-methyl vinyl isocyanate, idapangidwa ndikugwiritsidwa ntchito kutulutsa tinthu ta silika kuti tikonzekere polar ophatikizidwa stationary phase. makina kugwedera kuonetsetsa kuti homogeneous bedi aumbike mkati mwa column.Valuate odzaza ndime kulekana kwa peptide osakaniza wopangidwa asanu peptides;(Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) ndi Trypsin digest ya human serum albumin (HAS) .Kusakaniza kwa peptide ndi trypsin digest ya HSA inawonedwa kuti ilekanitse ndi kusamvana kwabwino komanso kothandiza. ma peptide ndi mapuloteni adawonedwa kuti adathetsedwa bwino komanso ogwira mtima pazanja la PMP, lomwe linali lothandiza kwambiri kuposa gawo la Ascentis Express RP-Amide.
PEG (Polyethylene Glycol), Urea, Acetic Acid, Trimethoxy Orthosilicate (TMOS), Trimethyl Chlorosilane (TMCS), Trypsin, Human Serum Albumin (HSA), Ammonium Chloride, Urea, Hexane Methyldisilazane (HMDS), Methacryloyl Chloride, BYPOYDORD (MC), Methacryloyl Chloride, BY-POHYD (MC) HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, ndi Acetone Yogulidwa kuchokera ku Sigma-Aldrich (St. Louis, MO, USA).
Chisakanizo cha urea (8 g), polyethylene glycol (8 g), ndi 8 mL wa 0.01 N acetic acid chinalimbikitsidwa kwa mphindi 10, ndiyeno 24 mL wa TMOS anawonjezeredwa pamenepo pansi pa nyengo yozizira kwambiri. 70 ° C kwa maola 12. Zouma zofewa zouma zinali zosalala pansi mu ng'anjo ndi calcined pa 550 ° C kwa maola 12. Magulu atatu adakonzedwa ndipo amadziwika kuti ayang'ane kuberekana mu kukula kwa tinthu, kukula kwa pore ndi pamwamba.
Pogwiritsa ntchito kusinthidwa kwa silika particles ndi pre-synthesized ligand phenylmaleimide-methylvinylisocyanate (PCMP) yotsatiridwa ndi ma polima a radial ndi styrene, gulu la polar lomwe lili ndi gulu linakonzedwa.Zosasunthika gawo la ma aggregates ndi unyolo wa polystyrene. Njira yokonzekera ikufotokozedwa pansipa.
N-phenylmaleimide (200 mg) ndi methyl vinyl isocyanate (100 mg) anasungunuka mu toluene youma, ndipo 0.1 mL ya 2,2'-azoisobutyronitrile (AIBN) anawonjezedwa mu botolo la reaction kuti akonze phenylmaleimide-methyl vinyl isocyanate copolymer wowuma pa 6 ° wosakaniza ndi fyuluta ya C3 ° yowumitsidwa mu 3 PM ° ndi fyuluta yowuma pa 6 ° C. uvuni pa 40 ° C kwa maola 3.
Zouma silika particles (2 g) anali omwazika mu youma toluene (100 mL), analimbikitsa ndi sonicated mu 500 mL wozungulira pansi botolo kwa 10 min.PMCP (10 mg) anali kusungunuka toluene ndipo anawonjezera dropwise kuti anachita botolo kudzera dontho funnel.The osakaniza anali refluxed pa 100 ° C zouma ndi zouma 100 ° C ndi zouma maola 100 ° C ndi zouma 100 ° C ndi zouma 100 ° C. maola.Kenako, PMCP-bonded silica particles (100 g) inasungunuka mu toluene (200 ml) ndi 4-hydroxy-TEMPO (2 mL) inawonjezeredwa pamaso pa 100 µL ya dibutyltin dilaurate monga chothandizira.Chisakanizocho chinagwedezeka pa 50 ° C ndi zouma kwa maola 8 °, zosefedwa kwa maola 8.
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), ndi TEMPO-PMCP-attached silica particles (1.5 g) anamwazikana mu toluene ndi kutsukidwa ndi nayitrogeni.The polymerization wa styrene unkachitika pa 100 ° C kwa maola 12. The chifukwa mankhwala anatsukidwa ndi methanol 60 ° C usiku zonse zouma chiwembu.
Zitsanzozo zinachotsedwa pa 393 K kwa ola la 1 kuti zipeze kupanikizika kotsalira kosachepera 10-3 Torr.Kuchuluka kwa N2 kudsorbed pamtundu wa P / P0 = 0.99 kunagwiritsidwa ntchito kuti adziwe kuchuluka kwa pore.The morphology ya bare ndi ligand-bonded silica particles, mwachitsanzo, Tokyo electron tachid scanning Tokyo, Tokyo, mwachitsanzo, kufufuzidwa kwa microscopy Tokyo. (silica yopanda kanthu ndi tinthu tating'ono ta silika) tinayikidwa pa aluminiyamu ndime pogwiritsa ntchito zomatira carbon tepi.Gold anali yokutidwa pa zitsanzo pogwiritsa ntchito Q150T sputter coater, ndi 5 nm Au wosanjikiza anaikidwa pa zitsanzo.Izi bwino ndondomeko ntchito bwino pogwiritsa ntchito voltages otsika ndipo amapereka mbewu zabwino, ozizira sputtering, EA2 Eleron, USA sputtering, USA sputtering, EA2 Eleron. A Malvern (Worcestershire, UK) Mastersizer 2000 tinthu kukula analyzer ntchito kupeza tinthu kukula kugawa. Naked silika particles ndi ligand-boma silika particles (5 mg aliyense) anali omwazika mu 5 mL wa isopropanol, sonicated kwa mphindi 10, vortexed, ndi kusanthula kwa 5 mingra pa 5 mg wa silika. amachitidwa pa mlingo wa 5 °C pa mphindi pa kutentha osiyanasiyana 30 mpaka 800 °C.
Zitsulo zosapanga dzimbiri zokhala ndi mizere yagalasi zokhala ndi mizere yopapatiza (id 100 × 1.8 mm) zidapakidwa pogwiritsa ntchito njira yolongedza slurry, kugwiritsa ntchito njira yomweyi yomwe imagwiritsidwa ntchito mu Ref.31.Chigawo chachitsulo chosapanga dzimbiri (chokhala ndi magalasi, 100 × 1.8 mm id) chokhala ndi chotuluka chokhala ndi 1 µm frit chinalumikizidwa ndi slurry packer (Alltech Deerfield, IL, USA). Konzani slurry ya gawo loyima poyimitsa 150 mg ya gawo lokhazikika la mL ndipo tumizani gawo lokhazikika la menotha mu menotha 1. slurry zosungunulira komanso propelling zosungunulira.Tsitsani ndime sequentially ntchito mavuto a MP 100 kwa mphindi 10, 80 MP kwa mphindi 15, ndi 60 MP kwa mphindi 30. Pakulongedza, makina kugwedera anagwiritsidwa ntchito ndi awiri GC ndime shakers (Alltech, Deerfield, IL kunyamula yunifolomu kumasula ndi kumasula yunifolomu kutulutsa pang'onopang'ono kutulutsa yunifolomu. ndime.Chotsani mzati kuchokera ku slurry wazolongedza unit ndikulumikiza koyenera kwina kolowera ndi ku dongosolo la LC kuti muwone momwe ntchito yake ikuyendera.
Pampu ya LC (10AD Shimadzu, Japan), injector (Valco (USA) C14 W.05) yokhala ndi 50nL jakisoni loop, membrane degasser (Shimadzu DGU-14A), UV-VIS capillary window inapangidwa Special µLC device detector (UV-2075) ndi magalasi olumikiza machubu aang'ono ang'onoang'ono aang'ono aang'ono aang'ono ndi magalasi aang'ono owonjezera. Ening.Atatha kulongedza, ma capillaries (50 μm id 365 ndi kuchepetsa ma capillaries (50 μm) anaikidwa pa 1/16 ″ kotuluka kwa mgwirizano wochepetsera. Northampton, MA).
Albumin yochokera ku seramu ya anthu, lyophilized ufa, ≥ 96% (agarose gel electrophoresis) 3 mg wothira trypsin (1.5 mg), 4.0 M urea (1 mL), ndi 0.2 M ammonium bicarbonate (1 mL) . The yankho anasonkhezeredwa kwa mphindi 10 ndi kusungidwa mu osamba madzi kwa 1 m 1 % ndi 60 FA osamba, ° C ndi 60 osamba T que 37. Sefa yankho ndikusunga pansi pa 4 °C.
Kupatukana kwa zosakaniza za peptide ndi ma HSA trypsin digests adawunikidwa padera pazaza za PMP. Onani kulekanitsidwa kwa peptide osakaniza ndi trypsin digest ya HSA ndi ndime ya PMP ndikuyerekeza zotsatira ndi gawo la Ascentis Express RP-Amide.
Zithunzi za SEM za tinthu ting'onoting'ono ta silika ndi tinthu tating'ono ta silika timawonetsedwa mu FIG.2 .Sem Zithunzi za tinthu tating'ono (A, b) zikuwonetsa izi, mosiyana ndi maphunziro athu akale, tinthu tating'onoting'ono tomwe timakhala ndi zingwe zosakhazikika.
Kusanthula zithunzi za maikulosikopu za ma elekitironi za tinthu tating'ono ta silika (A, B) ndi tinthu tating'ono ta silika (C, D).
The tinthu kukula magawidwe anabala silika particles ndi ligand-boma silika particles akuwonetsedwa Chithunzi 3 (A) .Volume-based particle size kugawa zokhotakhota anasonyeza kuti kukula kwa silika particles chinawonjezeka pambuyo mankhwala kusinthidwa (mkuyu 3A) .The tinthu kukula kugawa deta ya particles silika kuchokera panopa kuphunzira ndi phunziro lapitalo buku la particle1 (The particle-based 5). 3.36 μm, poyerekeza ndi phunziro lathu lapitalo ndi malonda (0.5) mtengo wa 3.05 μm (polystyrene-womangidwa silika particles) 34. Gululi linali ndi tinthu tating'onoting'ono tating'onoting'ono tating'onoting'ono poyerekezera ndi phunziro lathu lapitalo chifukwa cha kusiyana kwa PEG, urea, TMOS, ndi acetic acid mu zomwe timachita kusakaniza kwa polystyrene gawo lalikulu la particles . .Izi zikutanthauza kuti pamwamba functionalization wa silika particles ndi styrene yekha waikamo polystyrene wosanjikiza (0.97 µm) pa silika pamwamba, pamene mu gawo PMP wosanjikiza makulidwe anali 1.38 µm.
Tinthu kukula kugawa (A) ndi pore kukula kugawa (B) anabala silika particles ndi ligand-womangidwa silica particles.
Kukula kwa pore, pore volume ndi pamwamba pa silika particles ya kafukufuku wamakono amaperekedwa mu Table 1 (B) .The PSD mbiri za bare silica particles ndi ligand-bonded silica particles akuwonetsedwa mu Chithunzi 3 (B) Zotsatirazi zikufanana ndi phunziro lathu lapitalo. kukula kumachepa ndi 69 pambuyo pa kusintha kwa mankhwala, monga momwe tawonetsera mu Table 1 (B), ndipo kusintha kwa mphutsi kumasonyezedwa mu Chithunzi 3 (B) . Mofananamo, pore voliyumu ya silika particles inatsika kuchokera ku 0.67 mpaka 0.58 cm3 / g pambuyo pa kusintha kwa mankhwala. .Monga momwe tawonetsera mu Table 1 (B), pamtunda (m2 / g) wa silika particles adatsikanso kuchokera ku 116 m2 / g mpaka 105 m2 / g pambuyo pa kusintha kwa mankhwala.
Zotsatira za kusanthula kwapang'onopang'ono kwa gawo loyima zikuwonetsedwa mu Table 2. The carbon loading of panopa stationary phase ndi 6.35%, yomwe ili yotsika kusiyana ndi carbon loading ya phunziro lathu lapitalo (polystyrene bonded silica particles, 7.93% 35 ndi 10.21%, motsatira) 42. The carbon loading of the current stationary is 6.35%, yomwe ndi yotsika kusiyana ndi carbon loading ya phunziro lathu lapitalo (polystyrene bonded silica particles, 7.93% 35 ndi 10.21%, motsatira) 42. The carbon loading of the current stationary in the low, SP , polar s, polar, polar, SP, polar nd monga phenylmaleimide-methylvinylisocyanate (PCMP) ndi 4-hydroxy-TEMPO anagwiritsidwa ntchito.Kulemera kwa nayitrogeni peresenti ya gawo laposachedwa la stationary ndi 2.21%, poyerekeza ndi 0.1735 ndi 0.85% ndi kulemera kwa nayitrogeni m'maphunziro apitalo, motsatira. 4) ndi (5) anali 2.7% ndi 2.9%, motero, pamene kutulutsa mpweya wa mankhwala omaliza (6) anali 6.35%, monga momwe tawonetsera mu Table 2. Kutaya kulemera kunayang'aniridwa ndi PMP stationary phase, ndipo TGA curve ikuwonetsedwa mu Chithunzi 4. Mzere wa TGA umasonyeza kuchepa kwa kulemera kwa 3% yomwe ili ndi 8.6% yokhala ndi carbon, koma ndi C 8.6 yokhayo yomwe ili ndi 8.6% ya kaboni ndi yabwino. komanso N, O, ndi H.
The phenylmaleimide-methylvinylisocyanate ligand anasankhidwa kuti pamwamba kusinthidwa silika particles chifukwa ali polar phenylmaleimide magulu ndi magulu vinylisocyanate. mide moiety alibe ndalama pafupifupi pa pH yachibadwa. The polarity ya stationary gawo akhoza kulamulidwa ndi mulingo woyenera kuchuluka kwa styrene ndi nthawi anachita ya free radical polymerization. sitepe yotsiriza ya zimene anachita (free-radical polymerization) n'kofunika kwambiri ndipo akhoza kusintha polarity wa stationary gawo. mosemphanitsa.SPs okonzedwa ndi ndende zosiyanasiyana za styrene ndi osiyana carbon loadings.Apanso, katundu zoyima magawo mu mizati zosapanga dzimbiri zitsulo ndi fufuzani ntchito yawo chromatographic (selectivity, kusamvana, N mtengo, etc.) .Kutengera zoyesererazi, kukonzedwa wokometsedwa anasankhidwa kukonzekera PMP stationary gawo kuonetsetsa olamulira polarity ndi kusanthula bwino anate.
Zosakaniza zisanu za peptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) adayesedwanso pogwiritsa ntchito gawo la PMP pogwiritsa ntchito gawo la mafoni;60/40 (v/v) acetonitrile/madzi (0.1% TFA) pamlingo wothamanga wa 80 μL/mphindi. Pansi pa mikhalidwe yabwino kwambiri, nambala yambale yowerengera (N) pagawo lililonse (ID 100 × 1.8 mm) ndi 20,000 ± 100 (mbale 200,000 za NMP za P ndi 0000000). ma chromatogram akuwonetsedwa mu Chithunzi 5A.Kusanthula mwachangu pagawo la PMP pamlingo wothamanga kwambiri (700 μL/mphindi), ma peptides asanu adachotsedwa mkati mwa mphindi imodzi, ma N values ​​anali abwino kwambiri, 13,500 ± 330 pagawo lililonse (100 × 1.8 mm id), Amafanana ndi 0guden 135m0, 0. (100 × 1.8 mm id) anali odzaza ndi maere atatu osiyana a PMP yoyima gawo kuti aone reproducibility.The analyte ndende pa ndime iliyonse inalembedwa ntchito mulingo woyenera elution mikhalidwe ndi chiwerengero cha ongolankhula mbale N ndi posungira nthawi kulekanitsa chisakanizo chomwecho mayeso pa ndime.The reproducibility deta kwa PMPble ndende ndi mizati PMP bwino otsika ndime 4 ndime ya PMP otsika reproducilate ya PMP re. Mtengo wa RSD, monga momwe tawonetsera mu Gulu 3.
Kupatukana kwa peptide osakaniza pa PMP ndime (B) ndi Ascentis Express RP-Amide ndime (A);mobile phase 60/40 ACN/H2O (TFA 0.1%), PMP mizere mizere (100 × 1.8 mm id);analytical The elution dongosolo la mankhwala: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) ndi 5 (leucine) asidi enkephalin)).
Phokoso la PMP (100 × 1.8 mm ID) idayesedwa kuti iletse kukula kwa ma serratography albinography.Zakulu kwambiri Mu chromatogram (Chithunzi 6), chimbudzi cha HSA chagawika m'matumbo 17 a Peptides.
Kugaya kwa tryptic kwa HSA (100 × 1.8 mm id) kunapatulidwa pagawo la PMP;kuthamanga (100 µL/mphindi), gawo la m'manja 60/40 acetonitrile/madzi ndi 0.1% TFA.
kumene L ndi kutalika kwa mzere, η ndi kukhuthala kwa gawo la mafoni, ΔP ndi mzere wobwerera kumbuyo, ndipo u ndi liwiro laling'ono la gawo la mafoni. inali yofanana ndi ya phunziro lathu lapitalo Ref.34.The permeability wa mzati odzaza ndi mwachiphamaso porous particles ndi: 1.7 × 10-15 kwa 1.3 μm tinthu tating'onoting'ono, 3.1 × 10-15 kwa 1.7 μm particles, 5.2 × 10-5 × 2 μm 5 × 2 tinthu 2 μm particles 43. Choncho, kutsekemera kwa gawo la PMP kuli kofanana ndi 5 μm core-shell particles.
kumene Wx ndi kulemera kwa mzati wodzaza ndi chloroform, Wy ndi kulemera kwa mzati odzaza ndi methanol, ndi ρ ndi kachulukidwe zosungunulira. Kachulukidwe wa methanol (ρ = 0.7866) ndi chloroform (ρ = 1.484).The okwana porosity wa SILICA PARTICLES 108 × 18 × 18 × 18 × 1 8 × 1. Mizati ya Urea 31 yomwe tidaphunzira kale inali 0.63 ndi 0.55, motsatana. Izi zikutanthauza kuti kukhalapo kwa urea ligands kumachepetsa permeability ya gawo loyima. Komano, porosity yonse ya gawo la PMP (100 × 1.8 mm id) ndi 0.60.The permeability yodzaza ndi C1 particles mu CMP 1. Magawo 8 amtundu wa C18 ligands amamangiriridwa ku tinthu tating'onoting'ono ta silika ngati maunyolo ozungulira, pomwe m'magawo oyima amtundu wa polystyrene, wosanjikiza wokhuthala wa polymer umapangidwa mozungulira.
Chithunzi 7A,B chikuwonetsa ndime ya PMP (100 × 1.8 mm id) ndi Ascentis Express RP-Amide column (100 × 1.8 mm id) pogwiritsa ntchito mikhalidwe yofananira (ie, 60/40 ACN/H2O ndi 0.1% TFA).) ya chiwembu cha van Deemter.Zosakaniza zosankhidwa za peptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) zinakonzedwa mu 20 µL/ Kutsika kochepa kwa mizati yonse ndi 800 µL/min. ndi Express RP-Amide column anali 2.6 µm ndi 3.9 µm, motero.Makhalidwe a HETP amasonyeza kuti kulekanitsa bwino kwa gawo la PMP (100 × 1.8 mm id) ndikwabwino kwambiri kuposa malonda a Ascentis Express RP-Amide column (100 × 1.8 van dem id. osakhala ofunikira poyerekeza ndi phunziro lathu lapitalo.Kupambana kwakukulu kolekanitsa kwa gawo la PMP (100 × 1.8 mm id) poyerekeza ndi gawo la Ascentis Express RP-Amide limachokera ku kusintha kwa tinthu tating'onoting'ono, kukula kwake, ndi ndondomeko zovuta zonyamula mizere zomwe zimagwiritsidwa ntchito panopa34.
(A) van Deemter plot (HETP versus mobile phase linear velocity) yopezedwa pogwiritsa ntchito ndondomeko ya PMP (100 × 1.8 mm id) mu 60/40 ACN/H2O yokhala ndi 0.1% TFA.(B) van Deemter plot (HETP versus mobile phase linear velocity) yopezedwa pogwiritsa ntchito Ascent 6 mm 1 × 1 RP-Ascentis Express 0 ACN/H2O yokhala ndi 0.1% TFA.
Gawo lokhazikika la polar-embedded polystyrene stationary linakonzedwa ndikuwunikidwa kuti lilekanitse zosakaniza za peptide zopangidwa ndi trypsin ndi trypsin digests ya human serum albumin (HAS) mu chromatography yamadzimadzi yapamwamba kwambiri. particles, ankalamulira kaphatikizidwe wa stationary gawo, ndi zovuta mzati kulongedza katundu. Kuwonjezera mkulu kulekana dzuwa, otsika ndime kumbuyo kuthamanga pa otaya mitengo mkulu otaya ndi mwayi wina wa stationary phase.PMP mizati amasonyeza reproducibility wabwino ndipo angagwiritsidwe ntchito kusanthula peptide zosakaniza ndi trypsin chimbudzi zosiyanasiyana mapuloteni. ography.M'tsogolomu, mizati ya PMP idzawunikidwanso pa kulekanitsa kwa mapuloteni ndi ma antibodies a monoclonal.
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