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A na-akwadebe ihe ndị dị na silica porous site na usoro sol-gel na ụfọdụ mgbanwe iji nweta ihe ndị dị na macroporous. A na-emepụta ihe ndị a site na mgbakwunye na-atụgharị fragmentation chain transfer (RAFT) polymerization na N-phenylmaleimide-methylvinylisocyanate (PMI) na styrene iji kwadebe N-phenylmaleimstyrerrow steel-polystyrerrow (PMI). 100 × 1.8 mm id) juru n'ọnụ site na slurry packing. Nyochaa PMP kọlụm nkewa nke ngwakọta peptide mejupụtara ise peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin album) na-arụ ọrụ (Gly-Tyr, Gly-Leu-Tyr). Ọnọdụ na-adịghị mma, ọnụ ọgụgụ usoro ihe ọmụmụ nke ngwakọta peptide dị elu dị ka 280,000 efere / m². N'ịtụle ọrụ nkewa nke kọlụm mepere emepe na kọlụm Ascentis Express RP-Amide nke azụmahịa, a chọpụtara na arụmọrụ nkewa nke kọlụm PMP dị elu karịa kọlụm azụmahịa n'ihe gbasara arụmọrụ na mkpebi nkewa.
N'ime afọ ndị na-adịbeghị anya, ụlọ ọrụ biopharmaceutical abụrụla ahịa zuru ụwa ọnụ na-abawanye ụba na oke ahịa ahịa. Site na uto mgbawa nke ụlọ ọrụ biopharmaceutical1,2,3, a na-achọ nyocha nke peptides na proteins nke ukwuu. Na mgbakwunye na peptide lekwasịrị anya, a na-emepụta ọtụtụ adịghị ọcha n'oge njikọta peptide, si otú a na-achọ nyocha nke chromat na-achọ ka ọ dị ọcha. mmiri, anụ ahụ na sel bụ ọrụ siri ike nke ukwuu n'ihi ọnụ ọgụgụ dị ukwuu nke ụdị ndị a na-achọpụta na otu sample. Ọ bụ ezie na spectrometry mass bụ ngwá ọrụ dị irè maka peptide na protein sequencing, ma ọ bụrụ na a na-etinye ihe ndị dị otú ahụ n'ime spectrometer mass na otu ngafe, nkewa ahụ agaghị adị mma. Nke a nwere ike imebi nsogbu LC site n'imejuputa chromatography nke mmiri mmiri, nke ga-eme ka nyocha nke chromatography dị tupu ya abanye n'ime chromatography. spectrometer na oge a nyere4,5,6. Tụkwasị na nke ahụ, n'oge nkewa mmiri mmiri, ndị nyocha nwere ike ilekwasị anya na mpaghara ndị dị warara, si otú ahụ na-etinye uche na nyocha ndị a ma na-eme ka nchọpụta nchọpụta MS dịkwuo mma. Liquid chromatography (LC) abawanyela nke ukwuu n'ime afọ iri gara aga ma ghọọ usoro na-ewu ewu na proteomic analysis7,8,9,10.
Reversed-phase liquid chromatography (RP-LC) na-ọtụtụ ebe na-eji maka ịdị ọcha na nkewa nke peptide ngwakọta iji octadecyl-modified silica (ODS) dị ka adade phase11,12,13.However, RP guzo n'usoro adịghị enye afọ ojuju nkewa nke peptides na proteins n'ihi na ha mgbagwoju anya na-adọ 111,12,13. achọrọ iji nyochaa peptides na ndị na-edozi ahụ na polar na ndị na-abụghị polar moieties na-emekọrịta ma na-ejigide ndị a nyocha16.Mixed-mode chromatography, nke na-enye multimodal interactions, nwere ike ịbụ ihe ọzọ na RP-LC maka nkewa nke peptides, proteins, na ndị ọzọ mgbagwoju mix. Ọtụtụ mix-mode guzosie ike na kọlụm 1 a kwadebere na protein a kwadebere na separt usoro. 8,19,20,21.Mixed-mode guzo n'ihu (WAX / RPLC, HILIC / RPLC, polar intercalation / RPLC) na-adabara peptide na protein separations n'ihi ọnụnọ nke ma polar na ndị na-abụghị polar groups22,23,24,25,26,27,28 .N'otu aka ahụ na ike pụrụ iche na-egosi intervalating ọdụ ụgbọ okporo ígwè dị iche iche. ivity maka nyocha polar na ndị na-abụghị polar, dịka nkewa na-adabere na mmekọrịta dị n'etiti nyocha na oge nkwụsị.Multimodal mmekọrịta 29, 30, 31, 32. N'oge na-adịbeghị anya, Zhang et al.30 kwadebere a dodecyl-akwụsị polyamine ọdụ na-aga nke ọma kewapụrụ hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, na ọtụtụ ndị ọzọ analytes. The polar intercalator nwere ma polar na ndị na-abụghị polar otu, n'ihi ya, e nwere ike iji kewaa peptides na protein ndị nwere ma hydrophobic na hydrophilic-columbed-moieties, Ccomedded moieties. dị n'okpuru aha ahia Ascentis Express RP-Amide kọlụm, mana a na-eji kọlụm ndị a maka nyocha nke amine 33 naanị.
N'ime ọmụmụ ihe ugbu a, a kwadebere polar-embedded standary phase (N-phenylmaleimide-embedded polystyrene) ma nyochaa maka nkewa nke peptides na trypsin digests nke HSA. A na-akwado usoro nkwụsị ahụ site na iji usoro ndị a. A na-akwadebe ihe ndị dị na silica na-esi na ya dị ka usoro e nyere na mbipụta anyị gara aga na ụfọdụ mgbanwe nke glycol proglycol . OS, mmiri acetic acid na-edozi iji kwadebe ihe silica ahụ na nnukwu pore size. Nke abụọ, a ọhụrụ ligand, phenylmaleimide-methyl vinyl isocyanate, bụ synthesized na-eji na-derivatize silica particles iji kwadebe a polar agbakwunyere na-eguzosi ike phase.The n'ihi guzobe na-adọ e juru n'ọnụ n'ime a igwe anaghị agba nchara ngwugwu (1.8.8). na n'ibu vibration iji hụ na a homogeneous bed na-guzobere n'ime kọlụm. Nyochaa juru n'ọnụ kọlụm nkewa nke peptide ngwakọta mejupụtara ise peptides;(Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) na Trypsin digest nke mmadụ serum albumin (HAS) .The peptide ngwakọta na trypsin digest nke HSA e hụrụ ikewapụ na ezi mkpebi na arụmọrụ nke PRP bụ nke kọlụm tụnyere nke a sepa. A na-ahụ ma peptides na ndị na-edozi ahụ ka edozi ya nke ọma na ịrụ ọrụ nke ọma na kọlụm PMP, nke dị irè karịa kọlụm Ascentis Express RP-Amide.
PEG (Polyethylene Glycol), Urea, Acetic Acid, Trimethoxy Orthosilicate (TMOS), Trimetyl Chlorosilane (TMCS), Trypsin, Human Serum Albumin (HSA), Ammonium Chloride, Urea, Hexane Methyldisilazane (HMDS), Methacryloyl Chloride (Mc), Benzoroxide (Mc), Peroxide (Mc), Peroxide, Benzor, Peroxide, Benzor, Peroxide, Benzor. HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, na Acetone Zụrụ na Sigma-Aldrich (St. Louis, MO, USA).
Ngwakọta nke urea (8 g), polyethylene glycol (8 g), na 8 ml nke 0.01 N acetic acid na-akpali maka nkeji 10, wee tinye 24 mL nke TMOS n'ebe ahụ n'okpuru ọnọdụ oyi oyi. A na-ekpocha ngwakọta mmeghachi omume na 40 ° C maka awa 6 wee na 120 ° C na-agbanye mmiri na-ekpo ọkụ na mmiri na-adịghị na awa 7. 0 Celsius C maka awa 12. Ihe dị nro nke a mịrị amị bụ ala dị nro na oven na calcined na 550 ° C maka awa 12. A kwadebere batches atọ ma mara iji nyochaa reproducibility na nha nha, nha pore na ebe elu.
Site na ngbanwe elu nke silica particles na pre-synthesized ligand phenylmaleimide-methylvinylisocyanate (PCMP) na-esote radial polymerization na styrene, a kwadebere ngwakọta nwere otu polar.Oge nkwụsị maka nchịkọta na agbụ polystyrene. A kọwara usoro nkwadebe n'okpuru.
N-phenylmaleimide (200 mg) na methyl vinyl isocyanate (100 mg) gbazere na toluene akọrọ, na 0.1 ml nke 2,2′-azoisobutyronitrile (AIBN) agbakwunyere na flask mmeghachi omume iji kwadebe phenylmaleimide-methyl vinyl isocyanate copolymer na 0.1 ml nke akọrọ. oven na 40 Celsius C maka awa 3.
A na-ekesa ihe ndị na-esi na silica a mịrị amị (2 g) na toluene akọrọ (100 ml), kpaliri na sonicated na 500 mL gburugburu flask ala ala maka 10 min.PMCP (10 mg) na-agbaze na toluene ma gbakwunye dropwise na flask mmeghachi omume site na nkwụsịtụ funnel. A na-emegharị ngwakọta ahụ na 1800 C na-ekpo ọkụ maka awa 1800. 3 hours. Mgbe ahụ, a na-agbaze ihe ndị na-emepụta silica nke PMCP (100 g) na toluene (200 ml) na 4-hydroxy-TEMPO (2 mL) na-agbakwunye na ọnụnọ nke 100 µL nke dibutyltin dilaure dị ka ihe na-eme ka a na-eme ka ngwakọta ahụ na-agbanye na 50 ° C maka awa 8 na 5 ° C.
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), na TEMPO-PMCP na-ejikọta silica particles (1.5 g) na-agbasa na toluene ma kpochaa ya na nitrogen. A na-eme polymerization nke styrene na 100 ° C maka awa 12. A na-asacha ngwaahịa ahụ na methanol ° C n'ozuzu ya.
A na-ekpochapụ ihe nlele ahụ na 393 K maka awa 1 iji nweta nrụgide fọdụrụnụ nke na-erughị 10-3 Torr. Ọnụ ego nke N2 adsorbed na nrụgide ikwu nke P / P0 = 0.99 ejiri mee ihe iji chọpụta mkpokọta pore. ligand-bonded silica particles) na-etinye na kọlụm aluminom na-eji nrapado carbon teepu. A na-etinye ọla edo na ihe atụ na-eji Q150T sputter coater, na 5 nm Au oyi akwa na-edebe na samples. Nke a na-eme ka usoro nhazi nke ọma site na iji obere voltaji ma na-enye ọka dị mma, sputtering oyi. A Thermoti Electron, MA1 A na-eji ihe nyocha Flash 1. .A Malvern (Worcestershire, UK) Mastersizer 2000 particle size analyzer na-eji iji nweta urughuru nkesa nkesa.Naked silica particles na ligand-bonded silica particles (5 mg nke ọ bụla) na-agbasasị na 5 mL nke isopropanol, sonicated maka 10 min, vortexed maka 5 min, na a na-eme vortexed maka 5 min, na bekee. 5 Celsius C kwa nkeji n'elu oke okpomọkụ nke 30 ruo 800 Celsius.
Igwe anaghị agba nchara nwere kọlụm dị warara nke akụkụ (100 × 1.8 mm id) ejiri usoro nkwakọ ngwaahịa slurry, na-etinye otu usoro eji na Ref.31.A igwe anaghị agba nchara kọlụm (glass-lined, 100 × 1.8 mm id) na ntinye ntinye nke nwere 1 µm frit jikọtara ya na slurry packer (Alltech Deerfield, IL, USA) . Kwadebe slurry na-eguzo ọtọ site n'ịkwụsị 150 mg nke columnar mL na-ezigara ya na 1.Met. slurry ihe mgbaze nakwa dị ka propelling solvent. Jupụta kọlụm sequentially site n'itinye nrụgide nke 100 MP maka 10 nkeji, 80 MP maka 15 nkeji, na 60 MP maka 30 minutes. N'oge nkwakọ, n'ibu vibration e etinyere na abụọ GC kọlụm shakers (Alltech, Deerfield, IL, USA) iji hụ na kọlụm mebiri emebi n'ime kọlụm ọ bụla. .Kwụpụ kọlụm ahụ site na ngwugwu slurry ma jikọọ ọzọ dabara adaba na ntinye na na usoro LC iji lelee arụmọrụ ya.
Otu LC mgbapụta (10AD Shimadzu, Japan), injector (Valco (USA) C14 W.05) na 50nL injection loop, membrane degasser (Shimadzu DGU-14A), UV-VIS capillary window e wuru Pụrụ iche µLC ngwaọrụ detector (UV-2075) na iko-lined ejikọta microcolumns nke ukwuu. Mgbe nkwakọchara, a na-etinye capillaries (50 μm id 365 na mbenata otu capillaries (50 μm) na 1/16 ″ nke mbenata njikọ. A na-eme nchịkọta data na nhazi chromatographic site na iji sọftụwia Multichro 2000. A nwalere nlekota na 254 nm Oriọna Analytes maka UVginance data, MAAmpton Chromat.
Albumin sitere na ọbara mmadụ, ntụ ntụ lyophilized, ≥ 96% (agarose gel electrophoresis) 3 mg jikọtara ya na trypsin (1.5 mg), 4.0 M urea (1 mL), na 0.2 M ammonium bicarbonate (1 mL) . A na-emegharị ihe ngwọta maka nkeji 10 ma debe ya na mmiri ịsa ahụ na 37 ° C ruo awa 37.0 Celsius. Gbanwee ngwọta ma chekwaa n'okpuru 4 Celsius.
A na-enyocha nkewa nke ngwakọta peptide na HSA trypsin dị iche iche na kọlụm PMP. Lelee nkewa nke ngwakọta peptide na trypsin digest nke HSA site na kọlụm PMP ma tụnyere nsonaazụ ya na kọlụm Ascentis Express RP-Amide. A na-agbakọ ọnụ ọgụgụ efere usoro ihe omume dị ka ndị a:
Ihe onyonyo SEM nke ụmụ irighiri ihe silica efu na ụmụ irighiri silica nwere njikọ ligand ka egosiri na FIG.2 .Ma onyonyo nke ahihia dilica (a, b) gosiputara nke ahu, na akuku ndi ozo di iche iche, nke nwere ike ịbụ n'ihi mkpuchi nke ụdị polystyrene dị n'elu elu nke ndị silica.
Na-enyocha onyonyo microscope eletrọn nke ụmụ irighiri ihe silica efu (A, B) na ligand-bonded silica particles (C, D).
The urughuru size nkesa nke iferi silica ahụ na ligand-bonded silica ahụ na-egosi na Figure 3 (A) .Mpịakọta nke dabeere urughuru size nkesa curves gosiri na size nke silica ahụ mụbara mgbe chemical mgbanwe (Fig. 3A) The urughuru size nkesa data nke silica ahụ silica urughuru si ugbu a ọmụmụ na gara aga ọmụmụ bụ tụnyere .5.5. 3.36 μm, tụnyere anyị gara aga ọmụmụ na ad (0.5) uru nke 3.05 μm (polystyrene-bound silica particles)34.This ogbe nwere a narrower urughuru size nkesa tụnyere anyị gara aga ọmụmụ n'ihi na iche iche ruru nke PEG, urea, TMOS, na acetic acid na mmeghachi omume nke polystyre akụkụ nke ukwuu karịa na akụkụ nke si MP. Nke a pụtara na arụ ọrụ elu nke silica particles na styrene na-etinye naanị akwa polystyrene (0.97 µm) n'elu silica, ebe n'oge PMP, ọkpụrụkpụ oyi akwa bụ 1.38 µm.
Nkesa nha urughuru (A) na nkesa nha pore (B) nke ụmụ irighiri ihe silica na ihe silica na-ejikọta ligand.
The pore size, pore volume and surface area of ​​the silica particles of the current studies are given in Table 1 (B) .The PSD profaịlụ nke iferi silica particles na ligand-bonded silica particles e gosiri na Figure 3 (B) . Nsonaazụ dị tụnyere anyị gara aga ọmụmụ. The pore nha nke iferi na ligand0 na-egosi na silic 1 akụkụ nke 1 na-egosi na silic 1 akụkụ nke 3. re size na-ebelata site na 69 mgbe mgbanwe kemịkalụ, dị ka e gosiri na Isiokwu 1 (B), na mgbanwe nke curve na-egosi na Fig. 3 (B) .N'otu aka ahụ, pore olu nke silica ahụ mbelata 0.67 ka 0.58 cm3 / g mgbe chemical mgbanwe. The kpọmkwem elu ebe nke ugbu a na-amụ silica / gara aga bụ 2 particles (2 m 2) . g) .Dị ka e gosiri na Tebụl 1 (B), ebe elu (m2 / g) nke silica ahụ na-ebelata site na 116 m2 / g ruo 105 m2 / g mgbe mgbanwe kemịkalụ gasịrị.
The results nke elemental analysis nke guzo na-adọ na-egosi na Table 2. The carbon loading nke ugbu a guzo na-adọ bụ 6.35%, nke bụ ala karịa carbon loading nke anyị gara aga ọmụmụ (polystyrene bonded silica particles, 7.93% 35 na 10.21%, karị) 42. The carbon loading nke ugbu a na-adọ na-adọ na-adọ, n'ihi na ụfọdụ na-adọba n'ime SP. ligands dị ka phenylmaleimide-methylvinylisocyanate (PCMP) na 4-hydroxy-TEMPO e ji.The nitrogen arọ pasent nke ugbu a guzo na-adọ bụ 2.21%, tụnyere 0.1735 na 0.85% site arọ nke nitrogen na gara aga ọmụmụ, karị. Nke a pụtara na wt % nke carbonny ibu na-adọ n'ihi na nke ugbu a na-emepụta nitrogen dị elu n'ihi na nke na-emepụta ihe ndị na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta wt % nke carbonny na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-eme ka wt % nke carbonny na-emepụta ihe na-adabere na njedebe nke ọdụ ụgbọ mmiri. 4) na (5) bụ 2.7% na 2.9%, n'otu n'otu, mgbe carbon loading nke ikpeazụ ngwaahịa (6) bụ 6.35%, dị ka e gosiri na Tebụl 2. The arọ ọnwụ na-enyocha na PMP na-anọ ọdụ, na TGA curve na-egosi na Figure 4. The TGA curve na-egosi a arọ ọnwụ nke 8.6% carbon, ma na 3 n'ịdị mma n'ihi na 6.5 n'ihi na nkwekọ n'ihi na nkwekọ nke O. na H.
The phenylmaleimide-methylvinylisocyanate ligand a họọrọ maka elu mgbanwe nke silica ahụ n'ihi na ọ nwere polar phenylmaleimide òtù na vinylisocyanate ìgwè.Vinyl isocyanate iche iche nwere ike n'ihu meghachi omume na styrene site ndụ radical polymerization. The nke abụọ mere bụ itinye otu ìgwè nke nwere a agafeghị oke mmekọrịta na-adịghị ike na-adọ n'etiti analy na-adọ na analy. nylmaleimide moiety enweghị mebere ụgwọ na nkịtị pH.The polarity nke guzo na-adọ nwere ike na-achịkwa site na ezigbo ego nke styrene na mmeghachi omume oge nke free radical polymerization.The ikpeazụ nzọụkwụ nke mmeghachi omume (free-radical polymerization) dị oké egwu na nwere ike ịgbanwe polarity nke guzo phase.Elemental analysis e rụrụ iji lelee carbon na-aba ụba frasi nke ndị a na-eme nnyocha na-eme ka a na-elele nke carbon na-ebuwanye ibu nke ndị a ọdụ. Anọgide na-adọ na vice versa.SPs kwadebere na dị iche iche ịta nke styrene nwere dị iche iche carbon loadings.Again, Ibu ndị a guzo n'ụzọ n'ime igwe anaghị agba nchara ogidi ma lelee ha chromatographic arụmọrụ (selectivity, mkpebi, N uru, wdg) .Dabere na ndị a nnwale, ihe kachasị usoro a họọrọ iji kwadebe PMP guzo na-adọ iji hụ na-achịkwa polarity na ezi analyte.
A na-enyocha ngwakọta peptide ise (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) site na iji kọlụm PMP na-eji usoro mkpanaka;60/40 (v / v) acetonitrile / mmiri (0.1% TFA) na a eruba ọnụego nke 80 μL / min. N'okpuru ezigbo elution ọnọdụ, theoretical plate number (N) kwa kọlụm (100 × 1.8 mm id) bụ 20,000 ± 100 (200,000 na kọlụm nke chrome / m). A na-egosi atograms na eserese 5A. Nyocha ngwa ngwa na kọlụm PMP na oke ọsọ ọsọ (700 μL / min), peptides ise na-apụ n'anya n'ime otu nkeji, ụkpụrụ N dị ezigbo mma, 13,500 ± 330 kwa kọlụm (100 × 1.8 mm id), kwekọrọ na 135,000 plates / m (Fig 135,000). mm id) juru n'ọnụ atọ dị iche iche dị iche iche nke PMP ọdụ ọdụ iji lelee reproducibility. A na-edekọ mkpokọta nyocha maka kọlụm ọ bụla site na iji ọnọdụ elution kachasị mma na ọnụ ọgụgụ nke mbadamba ihe ọmụmụ N na oge njigide iji kewaa otu ngwakọta ule na kọlụm ọ bụla. A na-egosi data reproducibility maka ogidi PMP na Tebụl 4. A na-egosipụta nrụpụta nrụpụta nke ọma na kọlụm 3% dị na kọlụm PMPD.
Nkewa nke ngwakọta peptide na kọlụm PMP (B) na kọlụm Ascentis Express RP-Amide (A);Usoro mkpanaka 60/40 ACN/H2O (TFA 0.1%), akụkụ kọlụm PMP (100 × 1.8 mm id);analytical Usoro elution nke ogige: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) na 5 (leucine) acid enkephalin)).
A na-enyocha kọlụm PMP (100 × 1.8 mm id) maka nkewa nke tryptic digests nke ọbara albumin mmadụ na chromatography mmiri mmiri dị elu. chromatogram dị na Figure 6 na-egosi na ihe atụ ahụ kewapụrụ nke ọma na mkpebi ahụ dị mma nke ukwuu. A na-enyocha HSA digests site na iji ọsọ ọsọ nke 100 µL / min, mobile 0% egosi na TFA 70 µL / min. chromatogram (Njirimara 6), mgbaze HSA kewara n'ime ọnụ ọgụgụ 17 kwekọrọ na peptides 17. A na-agbakọ arụmọrụ nkewa nke ọ bụla na HSA digest na ụkpụrụ ndị a na-enye na tebụl 5.
A na-ekewapụta mgbaze tryptic nke HSA (100 × 1.8 mm id) na kọlụm PMP;Ọnụego nrịba (100 µL/min), usoro mkpanaka 60/40 acetonitrile/mmiri nwere 0.1% TFA.
ebe L bụ ogologo kọlụm, η bụ viscosity nke usoro mkpanaaka, ΔP bụ nrụgide azụ azụ, na u bụ ọsọ ọsọ ọsọ nke usoro mkpanaaka. The permeability nke PMP kọlụm bụ 2.5 × 10-14 m2, ọnụ ọgụgụ na-asọpụta bụ 25 μL / min, na 60/40 v / v ACN / mmiri dị ka 1 × 1.0. na nke anyị gara aga ọmụmụ Ref.34.The permeability nke kọlụm juru n'ọnụ na elu porous irighiri bụ: 1.7 × 10-15 maka 1.3 μm ahụ, 3.1 × 10-15 maka 1.7 μm ahụ, 5.2 × 10-15 na 2.5 × 10 μm akụkụ. 43.Ya mere, permeability nke usoro PMP yiri nke 5 μm core-shell particles.
ebe Wx bụ arọ nke kọlụm juru na chloroform, Wy bụ arọ nke kọlụm juru n'ọnụ na methanol, na ρ bụ njupụta nke ihe mgbaze.Densities nke methanol (ρ = 0.7866) na chloroform (ρ = 1.484) .The ngụkọta porosity nke SILICA PARTICLES 8 × 1 × 1 × 1 (3 mm) 8 × 1.0. rea ogidi 31 nke anyị na-amụ na mbụ bụ 0.63 na 0.55, karị. Nke a pụtara na ọnụnọ nke urea ligands na-ebelata permeability nke oge nkwụsị. N'aka nke ọzọ, ngụkọta porosity nke kọlụm PMP (100 × 1.8 mm id) bụ 0.60. The permeability nke PMP na-eme ka akụkụ nke CMP1 dị ala karịa akụkụ nke CMP1 nke CMP1 dị ala karịa akụkụ nke CMP1. -ụdị ọdụ ọdụ, C18 ligands na-ejikọta na silica particles dị ka linear chains, ebe na polystyrene-ụdị guzo n'usoro, A dịtụ oké polymer oyi akwa na-guzobere gburugburu ya. Na a-ahụkarị nnwale, na kọlụm porosity gbakọọ dị ka:
Ọgụgụ 7A, B na-egosi kọlụm PMP (100 × 1.8 mm id) na kọlụm Ascentis Express RP-Amide (100 × 1.8 mm id) na-eji otu ọnọdụ elution (ya bụ, 60/40 ACN / H2O na 0.1% TFA).) nke nkata van Deemter.A na-akwado ngwakọta peptide ahọpụtara (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) na 20 µL / Opekempe eruba maka kọlụm abụọ ahụ bụ 800 µL/min. Opekempe HETPm µL / min. bụ kọlụm Express RP-Amide bụ 2.6 µm na 3.9 µm, n'otu n'otu. Ụkpụrụ HETP na-egosi na nkewa nkewa nke kọlụm PMP (100 × 1.8 mm id) dị mma karịa ahịa dị na Ascentis Express RP-Amide (100 × 1.8 mm id) na-egosi ọnụ ahịa ọnụ ahịa nke ọnụ ahịa nke Ascentis Express RP-Amide (100 × 1.8 mm). Ọ bụghị ihe dị ịrịba ama ma e jiri ya tụnyere ọmụmụ ihe anyị gara aga. Ọrụ nkewa dị elu nke kọlụm PMP (100 × 1.8 mm id) ma e jiri ya tụnyere kọlụm Ascentis Express RP-Amide na-adabere na mmelite nke ọdịdị ahụ, nha, na usoro nchịkọta kọlụm dị mgbagwoju anya ejiri na ọrụ ugbu a34.
(A) van Deemter plot (HETP vesos mobile phase linear velocity) nwetara site na iji kọlụm PMP (100 × 1.8 mm id) na 60/40 ACN/H2O na 0.1% TFA. 0.1% TFA.
A kwadebere polar-agbakwunyere polystyrene ọdụ ọdụ ma nyochaa maka nkewa nke ngwakọta peptide synthetic na trypsin digests nke serum albumin mmadụ (HAS) na chromatography nke mmiri mmiri dị elu. a ahụ, achịkwa njikọ nke guzo na-adọ, na mgbagwoju kọlụm packing.In mgbakwunye na elu nkewa arụmọrụ, ala kọlụm azụ mgbali na elu eruba udu bụ ihe ọzọ uru nke a guzo phase.PMP ogidi exhibit ezi reproducibility na ike ga-eji maka analysis nke peptide ngwakọta na trypsin mgbaze nke dị iche iche proteins.We bu n'obi na-eji nke a na kọlụm nke na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe dị iche iche. chromatography.N'ọdịnihu, a ga-enyochakwa ogidi PMP maka nkewa nke protein na ọgwụ mgbochi monoclonal.
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