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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 ntinye ntinye nke mgbagwoju anya (RAFT) polymerization na N-phenylmaleimide-methylvinylisocyanate (PMI) na styrene iji kwadebe N-phenylmaleimstyrerrow nke ọdụ ụgbọ okporo ígwè (PMI). Ogidi igwe anaghị agba nchara (100 × 1.8 mm id) juru n'ọnụ site na slurry packing. Nyochaa nkewa kọlụm PMP nke ngwakọta peptide nke nwere peptides ise (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Argographic nke mmadụ na arụ ọrụ nke chromat chromat linfine) . serum albumin (HAS) .N'okpuru ọnọdụ elution kacha mma, ọnụ ọgụgụ usoro ihe ọmụmụ nke ngwakọta peptide dị elu dị ka 280,000 plates / m². N'iji ọrụ nkewa nke kọlụm mepere emepe na kọlụm Ascentis Express RP-Amide, a hụrụ na arụmọrụ nkewa nke kọlụm PMP dị elu karịa kọlụm nkewa na mkpebi nke effici.
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, yabụ na-achọ nyocha nke chromatization chọrọ. nke ndị na-edozi ahụ na mmiri mmiri, anụ ahụ na sel bụ ọrụ siri ike nke ukwuu n'ihi ọnụ ọgụgụ dị ukwuu nke ụdị ndị a na-achọpụta n'ime otu sample. Ọ bụ ezie na ọnụ ọgụgụ dị ukwuu nke MS bụ ngwá ọrụ dị irè maka peptide na protein sequencing, ma ọ bụrụ na a na-agbanye ihe ndị dị otú ahụ n'ime spectrometer mass na otu ngafe, nkewa ahụ agaghị adị mma. Nsogbu a nwere ike ibelata chromat nke chromat nke na-emejuputa atumatu nke chromographing tupu emejuputa ihe nyocha nke MS. nke ndị nyocha na-abanye na spectrometer mass na oge a na-enye4,5,6. Tụkwasị na nke ahụ, n'oge nkewa nke 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 analysis10.8,9.
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.Otú ọ dị, RP guzo n'ókè adịghị enye afọ ojuju nkewa nke peptides na proteins ruru ha mgbagwoju ọdịdị,1 amphire Ọdịdị na 1. A na-achọ usoro ndị a haziri ahazi iji nyochaa peptides na protein ndị nwere polar na ndị na-abụghị polar moieties na-emekọrịta ma na-ejigide chromatography ndị a16.Mixed-mode chromatography, nke na-enye multimodal interactions, nwere ike ịbụ ihe ọzọ na RP-LC maka nkewa nke peptides, protein, na ndị ọzọ mgbagwoju mix. Ọtụtụ kọlụm-mode-ejiwo na-adọ na-eji usoro ndị a. peptide na protein separations17,18,19,20,21.Mixed-mode guzo n'ókè (WAX / RPLC, HILIC / RPLC, polar intercalation / RPLC) na-adabara peptide na protein separations n'ihi ọnụnọ nke ma polar na ndị na-abụghị polar ìgwè22,23,24,25,282,282.7. covalently bonded pola iche iche na-egosi ezi nkewa ike na pụrụ iche selectivity maka pola na-abụghị polar analytes, dị ka nkewa dabere na mmekọrịta dị n'etiti nyocha na ọdụ na-adọ. Multimodal mmekọrịta 29, 30, 31, 32. N'oge na-adịbeghị anya, Zhang et al. 30 kwadebere usoro nkwụsị polyamine na-akwụsị dodecyl ma kewaa hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, na ọtụtụ nyocha ndị ọzọ. The polar intercalator nwere ma polar na ndị na-abụghị polar otu, n'ihi ya, a pụrụ iji ya kewaa peptides na protein ndị nwere ma hydrophobic na hydrophilic-coimbeded moieties. kọlụm) dị na azụmaahịa 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 ma nyochaa usoro nkwụsị nke polar (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 na-esi ísì ụtọ dị ka usoro e nyere na mbipụta anyị gara aga na ụfọdụ mgbanwe nke polyglycol proglycol. (PEG), TMOS, mmiri acetic acid e gbanwee iji kwadebe silica ahụ na nnukwu pore size.Nke abụọ, a ọhụrụ ligand, phenylmaleimide-methyl vinyl isocyanate, e synthesized na-eji na-derivatize silica ahụ iji kwadebe a pola agbakwunyere na-eguzosi ike n'ihe phase.The dapụtara na-eguzobe idless na-adọ na-eji ígwè na-eji igwe anaghị agba nchara 1 × 1.0. atụmatụ nkwakọ ngwaahịa. A na-enyere aka ịkwakọ kọlụm ahụ na vibration igwe iji hụ na e guzobere akwa akwa n'ime kọlụm. Nyochaa nkewa nke kọlụm nke ngwakọta peptide nke nwere peptides ise; (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) na Trypsin digest nke ụmụ mmadụ serum albumin (HAS) .The peptide ngwakọta na trypsin digest nke HSA hụrụ ikewapụ na ezi mkpebi na arụmọrụ nke PMP dị ka nke na-arụ ọrụ nke Express. Kọlụm RP-Amide. A na-ahụ ma peptides na ndị na-edozi ahụ na-edozi nke ọma na 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), Styroxy-H4, Benzor, Benzor, Styroxy-H4, Styroxy-H4H. Peroxide (BPO), 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 8. a mịrị amị na 70 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 na-akwadebe 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, 0.1 ml nke 2,2'-azoisobutyronitrile (AIBN). akọrọ na akpọnwụwo na oven na 40 Celsius 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 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 maka awa 180 na 180 ° C. 60 ° C maka awa 3. Mgbe ahụ, a na-agbaze ihe ndị na-emepụta silica nke PMCP (100 g) na toluene (200 ml) na 4-hydroxy-TEMPO (2 mL) n'ihu 100 µL nke dibutyltin dilaurate dị ka ihe na-akpali akpali. A na-emegharị ngwakọta ahụ na 80 ° C na awa 50 Celsius.
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 ka e ji mee ihe iji chọpụta mkpokọta pore. A na-etinye ya na kọlụm aluminom na-eji ihe ntanye 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 dị mma site na iji obere voltaji ma na-enye ọka dị mma, oyi spultham, USA Elelectron. A na-eji EA1112 elemental analyzer mee ihe maka nyocha nke elemental. A Malvern (Worcestershire, UK) Mastersizer 2000 particle size analyzer ka ejiri nweta nkesa nkesa. bench nke Mastersizer. A na-eme nyocha Thermogravimetric na ọnụego 5 Celsius C kwa nkeji karịa 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-eziga ya na 1 metan. e ji mee ihe dị ka slurry 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 hụ na kọlụm packing, IL, USA). nrụgide na-eji nwayọọ nwayọọ gbochie mmebi ọ bụla n'ime kọlụm ahụ. Kwụsị 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 enyo-lined njikọ obere obere tubing mmetụta. kọlụm band broadening.Mgbe nkwakọchara, capillaries (50 μm id 365 na mbenata union capillaries (50 μm) arụnyere na 1/16" ntinye nke mbenata n'otu. Data collection na chromatographic nhazi e mere site na iji Multichro 2000 software. Monitoring na 254 nm Analytes na-enyocha na 254 nm Analytes nke Chromat. (Northampton, MA).
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. 0.1% TFA.Nyochaa ngwọta na 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 .SEM ihe oyiyi nke ihe silica na-enweghị isi (A, B) na-egosi na, n'adịghị ka ọmụmụ ihe anyị gara aga, ihe ndị a na-eme ka ihe dị iche iche dị na nke a na-agbatị ma ọ bụ na-enwe ihe ngosi oge. ihe 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) .Volume 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 na-amụ na-tụnyere .5.5. nke PMP bụ 3.36 μm, tụnyere anyị gara aga ọmụmụ na ad (0.5) uru nke 3.05 μm (polystyrene-ejikọta silica ahụ) 34.This ogbe nwere a narrower urughuru size nkesa tụnyere anyị gara aga ọmụmụ ruru ka iche iche ruru nke PEG, urea, TMOS, na acetic acid na-adọ nke dịtụ ukwuu nke ngwakọta nke ngwakọta nke PEG. Nke a pụtara na n'elu functionalization nke silica ahụ na styrene naanị edebe polystyrene oyi akwa (0.97 µm) na silica elu, ebe na PMP na-adọ 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 profiles of iferi silica particles na ligand-bonded silica particles na-egosi na Figure 3 (B) . Nsonaazụ dị tụnyere anyị gara aga ọmụmụ. The pore nha nke iferi na ligand0 na silic 1 akụkụ nke 3, nke 1 silica na-asọpụrụ 1. na-egosi na nha pore na-ebelata site na 69 mgbe mgbanwe kemịkalụ gasịrị, dị ka e gosiri na Tebụl 1 (B), na mgbanwe nke ntụgharị ahụ na-egosi na Fig. 3 (B) .N'otu aka ahụ, ụda pore nke silica particles na-ebelata site na 0.67 ruo 0.58 cm3 / g mgbe mgbanwe kemịkalụ gasịrị. Ebe a kapịrị ọnụ n'elu akụkụ nke silic1 nke ugbu a bụ m2 nke silica na-amụ 1 m. ọmụmụ ihe anyị gara aga (124 m2 / g) .Dị ka e gosiri na Tebụl 1 (B), ebe dị elu (m2 / g) nke silica particles na-ebelatakwa site na 116 m2 / g ruo 105 m2 / g mgbe mgbanwe kemịkalụ gasịrị.
A na-egosi nsonaazụ nke elemental nyocha nke oge nkwụsị na tebụl 2. The carbon loading nke ugbu a na-anọ ọdụ bụ 6.35%, nke dị ala karịa carbon loading nke anyị gara aga ọmụmụ (polystyrene bonded silica particles, 7.93% 35 na 10.21%, n'otu n'otu) 42. The carbon loading nke ugbu a na-adọ na-adọ na-adọ na-adọ na-adọ, n'ihi na nke ugbu a nkwadebe nke SP dị ala. styrene, ụfọdụ polar ligands dị ka phenylmaleimide-methylvinylisocyanate (PCMP) na 4-hydroxy-TEMPO were used.The nitrogen arọ percent 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 nitrogen na-adọ na-adị ugbu a na-adọ na wt . phenylmaleimide.N'otu aka ahụ, carbon loadings nke ngwaahịa (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 guzo na-adọ, na TGA curve e gosiri na Figure 4. Ngwunye carbon (6.35%) n'ihi na ligands nwere ọ bụghị naanị C kamakwa N, 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 ìgwè nwere ike n'ihu meghachi omume na styrene site ndụ radical polymerization. The abụọ mere bụ itinye otu ìgwè nke nwere a agafeghị oke mmekọrịta na ọ dịghị eletrọnịkị mmekọrịta na analy ọdụ. na-adọ, ebe ọ bụ na phenylmaleimide moiety enweghị mebere ụgwọ na nkịtị pH.The polarity nke guzo na-adọ nwere ike ịchịkwa site 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 ada ada phase.Elemental analysis e rụrụ na-elele na-adọ na-adọ na-adọ nke carbon analysis e rụrụ na-elele na-adọ na-adọ. na mmeghachi omume oge ụba carbon loading nke guzo 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 na-elele ha chromatographic arụmọrụ (selectivity, mkpebi, N uru, wdg) .Dabere na ndị a nnwale, ihe kachasị usoro a họọrọ iji hụ na ezi PMP na-achịkwa ọdụ ọdụ na-eme ka a na-ahọrọ iji kwadebe PMP na-achịkwa ọdụ ọdụ.
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 ọsọ ọsọ 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 ² nke PPA / m atọ). A na-egosi ogidi na chromatograms na Figure 5A. Nyocha ngwa ngwa na kọlụm PMP na ọnụ ọgụgụ dị elu (700 μL / min), peptides ise na-agbapụta n'ime otu nkeji, ụkpụrụ N dị ezigbo mma, 13,500 ± 330 kwa kọlụm (100 × 1.8 mm id), kwekọrọ na 135 / 00 Fig). Ogidi (100 × 1.8 mm id) juru n'ọnụ atọ dị iche iche nke PMP na-anọ ọdụ iji lelee reproducibility. A na-edekọ mkpokọta nyocha maka kọlụm ọ bụla site na iji ọnọdụ elution kachasị mma na ọnụ ọgụgụ nke ihe ngosi ihe ngosi N na oge njigide iji kewaa otu ngwakọta ule na kọlụm ọ bụla. A na-egosipụta data reproducibility maka kọlụm PMP nke ọma na kọlụm PMP nke ọma na-egosipụta na kọlụm PMP nke ọma na kọlụm 4. %RSD ụkpụrụ, dị ka e gosiri na Tebụl 3.
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 nlele ahụ kewapụrụ nke ọma na mkpebi ahụ dị ezigbo mma. A na-enyocha HSA digests site na iji ọsọ ọsọ nke 100 µL / min, mobile phase 70 µL / min, mobile phase 70. TFA.Dị ka egosiri na chromatogram (Nyocha 6), e kewara mgbaze HSA 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 ngwa ngwa ngwa ngwa nke usoro mkpanaaka. The permeability nke kọlụm PMP bụ 2.5 × 10-14 m2, ọnụ ọgụgụ na-agba ọsọ bụ 25 μL / min, na 60/40 v / v ACN / mmiri na-eji 1 × 0. id) yiri 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 2.2 × 2.2 × 10-15 na 2.2 μm. ahụ Maka 5 μm particles 43. Ya mere, permeability nke oge PMP yiri nke 5 μm isi-shell ahụ.
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 1 × 1 × 1 × 1 (1 mm). Ogidi C18-Urea 31 nke anyị na-amụbu bụ 0.63 na 0.55, n'otu n'otu. Nke a pụtara na ọnụnọ nke urea ligands na-ebelata njedebe nke oge nkwụsị. N'aka nke ọzọ, ngụkọta porosity nke kọlụm PMP (100 × 1.8 mm id) bụ 0.60. The permeability nke columned dị ala karịa PMP1. silica particles n'ihi na n'ụdị C18-ụdị ọdụ ọdụ, C18 ligands na-ejikọta na silica particles dị ka linear chains, mgbe 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/ Ọnụego kacha nta maka kọlụm abụọ ahụ bụ 800 µL/min. Opekempe HETPm na ọnụego kacha nta (800 µL / min). kọlụm na kọlụm Ascentis Express RP-Amide bụ 2.6 µm na 3.9 µm, n'otu n'otu. Ụkpụrụ HETP na-egosi na arụmọrụ nkewa nke kọlụm PMP (100 × 1.8 mm id) dị mma karịa ahịa ahịa dị na Ascentis Express RP-Amide kọlụm (100 × 1. Det). na mbelata nke uru N na mmụba na-arịwanye elu abụghị 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 dabeere na mmelite na ọdịdị ahụ, nha, na usoro nchịkọta kọlụm dị mgbagwoju anya na-eji 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. ACN/H2O nwere 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 dị elu nke mmiri mmiri. size nke silica 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 na-edozi ahụ na-eji kọlụm nke a na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe na-emepụta ihe dị iche iche na-edozi ahụ. osisi ọgwụ na fungal extracts na chromatography mmiri mmiri.N'ọdịnihu, a ga-enyocha ogidi PMP maka nkewa nke protein na monoclonal antibodies.
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