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Ua hoʻomākaukau ʻia nā ʻāpana silica porous e kahi ʻano sol-gel me kekahi mau hoʻololi e loaʻa ai nā ʻāpana macroporous. (100 × 1.8 mm id) ua paʻi ʻia e ka slurry packing.Evaluated PMP kolamu hoʻokaʻawale ʻana o kahi hui peptide me ʻelima peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg) (HAS).Ma lalo o nā kūlana elution maikaʻi loa, ʻoi aku ka kiʻekiʻe o ka helu theoretical plate o ka hui peptide e like me 280,000 mau papa / m². Ke hoʻohālikelike ʻana i ka hana hoʻokaʻawale ʻana o ke kolamu i kūkulu ʻia me ka kolamu Ascentis Express RP-Amide kalepa, ua ʻike ʻia ʻoi aku ka maikaʻi o ka hoʻokaʻawale ʻana o ka kolamu PMP ma mua o ke kolamu kalepa ma ke ʻano o ka hoʻokaʻawale ʻana a me ka hoʻonā.
I nā makahiki i hala iho nei, ua lilo ka ʻoihana biopharmaceutical i mea hoʻonui i ka mākeke honua me ka piʻi nui ʻana o ka māhele mākeke. ʻO nā protein i loko o nā wai kino, nā ʻiʻo a me nā cell he hana paʻakikī loa ma muli o ka nui o nā ʻano mea hiki ke ʻike ʻia i loko o kahi laʻana hoʻokahi. analytes e komo ana i ka nuipa spectrometer i ka manawa i haawiia 4,5,6. Eia kekahi, i ka wā o ka hoʻokaʻawale ʻana o ka wai, hiki ke hoʻopaʻa ʻia nā analytes i nā ʻāpana liʻiliʻi, a laila e noʻonoʻo ana i kēia mau analytes a me ka hoʻomaikaʻi ʻana i ka sensitivity ʻike MS. Ua holomua nui ka chromatography wai (LC) i nā makahiki he ʻumi i hala a ua lilo i ʻenehana kaulana i ka proteomic, 17.
Hoʻohana nui ʻia ka reversed-phase liquid chromatography (RP-LC) no ka hoʻomaʻemaʻe ʻana a me ka hoʻokaʻawale ʻana o nā hui peptide me ka hoʻohana ʻana i ka silica octadecyl-modified (ODS) ma ke ʻano he stationary phase11,12,13. Akā naʻe, ʻaʻole hāʻawi nā RP stationary phases i ka hoʻokaʻawale maikaʻi ʻana o nā peptides a me nā protein ma muli o ko lākou ʻano paʻakikī 14, philore i hoʻolālā ʻia a me ka amphire. Pono nā māhele e kālailai i nā peptides a me nā protein me nā polar a me nā ʻāpana polar ʻole e launa pū me ka mālama ʻana i kēia mau analytes16. Mixed-mode chromatography, e hāʻawi ana i nā pilina multimodal, hiki ke lilo i mea ʻokoʻa i ka RP-LC no ka hoʻokaʻawale ʻana i nā peptides, proteins, a me nā hui paʻakikī ʻē aʻe. hoʻokaʻawale17,18,19,20,21.Mixed-mode stationary phases (WAX/RPLC, HILIC/RPLC, polar intercalation/RPLC) he mea kūpono no ka peptide a me ka hoʻokaʻawale protein ma muli o ka loaʻa ʻana o nā pūʻulu polar a me nā pūʻulu polar ʻole22,23,24,25,26,27,28 intercalation station, polarly,28. Hōʻike nā pūʻulu polar paʻa i ka mana hoʻokaʻawale maikaʻi a me ke koho kūʻokoʻa no nā analytes polar a me ka polar ʻole, no ka mea, pili ka kaʻawale ʻana i ka pilina ma waena o ka analyte a me ka pae paʻa. ʻO nā pilina multimodal 29, 30, 31, 32. ʻO Zhang et al. 30 hoʻomākaukau i ka dodecyl-terminated polyamine stationary phase a me ka hoʻokaʻawale maikaʻi i nā hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, a me kekahi mau analytes ʻē aʻe. ʻO ka intercalator polar he mau polar a me nā pūʻulu non-polar, no laila hiki ke hoʻohana ʻia e hoʻokaʻawale i nā peptides a me nā protein i loaʻa nā hydrophobic a me hydrophilic moieties. nā kolamu) i kūʻai ʻia ma lalo o ka inoa kālepa ʻo Ascentis Express RP-Amide kolamu, akā hoʻohana ʻia kēia mau kolamu no ka nānā ʻana i ka amine 33 wale nō.
Ma ka haʻawina o kēia manawa, ua hoʻomākaukauʻia a loiloiʻia kahi papa hana polar-embedded (N-phenylmaleimide-embedded polystyrene) no ka hoʻokaʻawaleʻana i nā peptides a me nā trypsin digests o HSA. Ua hoʻomākaukauʻia ka pae paʻa me ka hoʻohanaʻana i ka papahana. Ua hoʻoponopono ʻia ka waikawa no ka hoʻomākaukau ʻana i nā ʻāpana silica me ka nui pore nui. ʻO ka lua, kahi ligand hou, phenylmaleimide-methyl vinyl isocyanate, ua synthesized a hoʻohana ʻia e derivatize i nā ʻāpana silica no ka hoʻomākaukau ʻana i ka polar embedded stationary phase. e hōʻoia i kahi moe homogeneous i hoʻokumu ʻia i loko o ke kolamu. (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) a me Trypsin digest o ke kanaka serum albumin (HAS) . ʻO ke kolamu RP-Amide. Ua ʻike ʻia nā peptides a me nā protein i hoʻoholo maikaʻi ʻia a maikaʻi hoʻi ma ka kolamu PMP, ʻoi aku ka maikaʻi ma mua o ke kolamu 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 (MC-OTEMP), Stydroxy-Oxide (MC-OTEMP) HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, a me Acetone i kūʻai ʻia mai Sigma-Aldrich (St. Louis, MO, USA).
Ua hoʻoulu ʻia kahi hui ʻana o ka urea (8 g), polyethylene glycol (8 g), a me 8 mL o 0.01 N acetic acid no 10 mau minuke, a laila hoʻohui ʻia he 24 mL o TMOS ma lalo o nā kūlana hau. ua maloʻo ma 70 ° C no nā hola 12. ʻO ka mea maloʻo maloʻo maloʻo he lepo maʻemaʻe i loko o ka umu a calcined ma 550 ° C no nā hola 12. Ua hoʻomākaukauʻia nā'āpanaʻekolu a uaʻikeʻia e nānā i ka hana houʻana i ka nui o ka pore, ka nui o ka pore a me ka ili.
Ma ka hoʻololi ʻana i ka ʻili o nā ʻāpana silica me ka ligand pre-synthesized phenylmaleimide-methylvinylisocyanate (PCMP) i ukali ʻia e ka polymerization radial me ka styrene, ua hoʻomākaukau ʻia kahi hui pūʻulu polar. ʻO ka pae paʻa no nā aggregates a me nā kaulahao polystyrene. Ua wehewehe ʻia ke kaʻina hana hoʻomākaukau ma lalo nei.
N-phenylmaleimide (200 mg) a me methyl vinyl isocyanate (100 mg) ua hoʻoheheʻeʻia i loko o ka toluene maloʻo, a me 0.1 mL o 2,2′-azoisobutyronitrile (AIBN) i hoʻohuiʻia i ka pahu hopena e hoʻomākaukau ai i ka phenylmaleimide-methyl isocyanate copolymer (PMC) a me 3 mau hola i hoʻohuiʻia. maloʻo i loko o ka umu ma 40 ° C no 3 mau hola.
Ua hoʻopuehu ʻia nā ʻāpana silica maloʻo (2 g) i loko o ka toluene maloʻo (100 mL), hoʻoulu ʻia a sonicated i loko o kahi pahu 500 mL puni lalo no 10 min. PMCP (10 mg) ua hoʻoheheʻe ʻia i loko o toluene a hoʻohui ʻia i ka dropwise i ka pahu hopena ma o ka hāʻule ʻana o ka funnel. 60 ° C no nā hola 3. A laila, ua hoʻoheheʻe ʻia nā ʻāpana silica paʻa PMCP (100 g) i toluene (200 ml) a me 4-hydroxy-TEMPO (2 mL) i hoʻohui ʻia i mua o 100 µL o dibutyltin dilaurate ma ke ʻano he catalyst.
Ua hoʻopuehu ʻia ʻo Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), a me TEMPO-PMCP-attached silica particles (1.5 g) i toluene a hoʻomaʻemaʻe ʻia me ka nitrogen. Ua lawe ʻia ka polymerization o styrene ma 100 ° C no nā hola 12. Ua holoi ʻia ka huahana me ka methanol a maloʻo i ka pō.
Ua hoʻokuʻu ʻia nā mea hoʻohālike ma 393 K no 1 hola e loaʻa ai ke koena kaomi o ka liʻiliʻi ma lalo o 10-3 Torr. ʻO ka nui o N2 adsorbed ma kahi pili pili o P / P0 = 0.99 ua hoʻohana ʻia e hoʻoholo ai i ka nui o ka pore volume.The morphology of bare and ligand-bonded silica particles i nānā ʻia me ka scanning electron micros, Japan samples. a me nā ʻāpana silica i hoʻopaʻa ʻia i ligand) ua kau ʻia ma luna o ke kolamu alumini me ka hoʻohana ʻana i ka lipine carbon adhesive. Malvern (Worcestershire, UK) Mastersizer 2000 particle size analyzer ua hoʻohana 'ia no ka loaʻaʻana o ka nui particle distribution.Naked silica particles a me ka ligand-hoʻopaʻa silica particles (5 mg kēlā me kēia) ua hoopuehuia ma 5 mL o isopropanol, sonicated no 10 min, vortexed no 5 min, a kau ma luna o ka optical bench ma luna o ka 5 mau minuke i hoʻopaʻa 'ia ma luna o ka Optical bench o ka helu o Masters. ka wela wela o 30 a 800 °C.
Ua hoʻopili ʻia nā kolamu ololi o nā ana (100 × 1.8 mm id) me ka hoʻohana ʻana i ke ʻano slurry packing, e pili ana i ke kaʻina hana like i hoʻohana ʻia ma Ref. 31. Ua hoʻohui ʻia kahi kolamu kila kuhiliʻole (ke aniani-laina, 100 × 1.8 mm id) me kahi puka puka i loaʻa kahi frit 1 µm i hoʻopili ʻia i kahi packer slurry (Alltech Deerfield, IL, USA). slurry solvent a me ka propelling solvent. E hoʻopiha i ke kolamu sequentially ma ka noi ʻana i nā kaomi o 100 MP no 10 mau minuke, 80 MP no 15 mau minuke, a me 60 MP no 30 mau minuke. I ka wā o ka hoʻopaʻa ʻana, ua hoʻopili ʻia ka haʻalulu mechanical me ʻelua GC column shakers (Alltech, Deerfield, IL, USA) e hōʻoia i ka paʻi paʻi paʻi o ke kolamu. no ka pale ʻana i ka pōʻino i loko o ke kolamu. E hoʻokaʻawale i ke kolamu mai ka ʻāpana slurry packing a hoʻohui i kekahi mea kūpono i ka puka komo a me ka ʻōnaehana LC e nānā i kāna hana.
He pamu LC (10AD Shimadzu, Iapana), injector (Valco (USA) C14 W.05) me 50nL injection loop, membrane degasser (Shimadzu DGU-14A), UV-VIS capillary window i kukuluia. broadening.Ma hope o ka hoʻopili ʻana, ua hoʻokomo ʻia nā capillaries (50 μm id 365 a me ka hōʻemi ʻana i nā capillaries union (50 μm) ma ka 1/16 ″ puka o ka hui hoʻohaʻahaʻa. Ua hana ʻia ka hōʻiliʻili ʻikepili a me ka hana chromatographic me ka hoʻohana ʻana i ka polokalamu Multichro 2000. ʻO ka nānā ʻana ma 254 nm Ua hoʻāʻo ʻia nā ʻikepili ʻo ChrodPro 8. (Northampton, MA).
Albumin mai ke kanaka serum, lyophilized pauka, ≥ 96% (agarose gel electrophoresis) 3 mg i hui me trypsin (1.5 mg), 4.0 M urea (1 mL), a me 0.2 M ammonium bicarbonate (1 mL). Ua hoʻonāukiʻia ka hopena no 10 mau minuke a mālamaʻia i loko o ka waiʻauʻau ma 37, 1 °C. 0.1% TFA. E kānana i ka hopena a mālama i lalo o 4 °C.
Hoʻokaʻawale ʻia ka hoʻokaʻawale ʻana o ka hui peptide a me ka HSA trypsin digests ma nā kolamu PMP.
Hōʻike ʻia nā kiʻi SEM o nā ʻāpana silika ʻole a me nā ʻāpana silika i hoʻopaʻa ʻia i ka ligand ma FIG. 2. Hōʻike nā kiʻi SEM o nā ʻāpana silica ʻole (A, B) i ka ʻokoʻa i kā mākou mau noiʻi mua ʻana, he spherical kēia mau ʻāpana kahi e hoʻolōʻihi ʻia ai nā mea liʻiliʻi a i ʻole like ʻole ke ʻano like ʻole.
Ke nānā nei i nā kiʻi microscope electron o nā ʻāpana silica ʻole (A, B) a me nā ʻāpana silica paʻa ligand (C, D).
Hōʻike ʻia ka puʻunaue ʻana o ka nui o nā ʻāpana silica a me nā mea liʻiliʻi i hoʻopaʻa ʻia ma ke kiʻi 3 (A). Hōʻike ʻia ka nui o ka nui o ka nui o nā silica ma hope o ka hoʻololi ʻana (Fig. 3A). μm, i hoʻohālikelike ʻia me kā mākou noiʻi mua ʻana me ka waiwai ad (0.5) o 3.05 μm (polystyrene-bound silica particles) 34. Ua ʻoi aku ka liʻiliʻi o kēia puʻupuʻu i ka nui o ka nui o ka nui ma mua o kā mākou noiʻi mua ʻana ma muli o nā ʻano like ʻole o ka PEG, urea, TMOS, a me ka acetic acid i ka hui ʻana. ʻO ka hoʻohana ʻana i ka ili o nā ʻāpana silica me ka styrene i waiho wale i kahi papa polystyrene (0.97 µm) ma luna o ka ʻili silica, akā ma ka pae PMP he 1.38 µm ka mānoanoa o ka papa.
Ka puunaue ana i ka nui o ka palaki (A) a me ka puunaue nui o ka pore (B) o na mea silika olohelohe a me na kinikini silica paa ligand.
ʻO ka nui o ka pore, ka nui pore a me ka ʻili o nā ʻāpana silica o ka haʻawina o kēia manawa ua hāʻawi ʻia ma ka Papa 1 (B). ʻO nā kiʻi PSD o nā ʻāpana silica ʻole a me nā ʻāpana silica i hoʻopaʻa ʻia i ligand i hōʻike ʻia ma ke Kiʻi 3 (B). Hoʻohālikelike ʻia nā hopena i kā mākou noiʻi mua. e 69 ma hope o ka hoʻololi kemika, e like me ka mea i hōʻike ʻia ma ka Papa 1(B), a ua hōʻike ʻia ka hoʻololi ʻana o ka pihi ma Fig. 3(B) .Pēlā, ua emi ka nui o ka pore o nā silica particles mai 0.67 a i 0.58 cm3/g ma hope o ka hoʻololi kemika. ʻO ka ʻāpana kiko o ka ʻāpana silica m2 i aʻo ʻia i kēia manawa he 116 (ʻo kā mākou mau mea silica m2 i aʻo ʻia i kēia manawa he 116. m2/g). E like me ka mea i hoikeia ma ka Papa 1(B), ua emi iho ka ili (m2/g) o na mea silica mai 116 m2/g a hiki i 105 m2/g ma hope o ka hoololi kemika.
Hōʻike ʻia nā hopena o ka loiloi elemental o ka pae paʻa ma ka Papa 2. ʻO ka hoʻouka kalapona o ka manawa paʻa o kēia manawa he 6.35%, ʻoi aku ka haʻahaʻa ma mua o ka hoʻouka kalapona o kā mākou haʻawina mua (polystyrene bonded silica particles, 7.93%35 a me 10.21%, kēlā me kēia) 42. ʻO ka hoʻouka kalapona o ka SP stationary phase o kēia manawa he haʻahaʻa, no ka mea, haʻahaʻa ka hoʻouka kalapona o ka SP stationary i kēia manawa, no ka mea, haʻahaʻa ka hoʻouka ʻana o ke kalapona SP i kēia manawa, no ka mea, he haʻahaʻa ka hoʻouka kalapona o ka māhele paʻa SP i kēia manawa, no ka mea, haʻahaʻa ka poʻe. Ua hoʻohana ʻia nā ligands e like me phenylmaleimide-methylvinylisocyanate (PCMP) a me 4-hydroxy-TEMPO. ʻO ka pākēneka paona nitrogen o ka manawa paʻa o kēia manawa he 2.21%, i hoʻohālikelike ʻia me 0.1735 a me 0.85% ma ke kaumaha o ka nitrogen i nā haʻawina mua. ʻO ka hoʻouka ʻana o nā huahana (4) a me (5) he 2.7% a me 2.9%, ʻoiai ʻo ka hoʻouka ʻana o ke kalapona o ka huahana hope loa (6) he 6.35%, e like me ka hōʻike ʻana ma ka Papa 2. Ua nānā ʻia ka poho kaumaha me ka PMP stationary phase, a ua hōʻike ʻia ka TGA curve ma ke kiʻi 4. ʻO ka TGA curve e hōʻike ana i ka poho kaumaha o 8.6% no ka mea, he 8.6% maikaʻi. ʻAʻole ʻo C wale nō nā ligand akā ʻo N, O, a me H.
Ua koho ʻia ka ligand phenylmaleimide-methylvinylisocyanate no ka hoʻololi ʻana i ka ʻili o nā ʻāpana silica no ka mea he mau pūʻulu phenylmaleimide polar a me nā hui vinylisocyanate. Hiki i nā hui isocyanate Vinyl ke hana hou aku me ka styrene e ka polymerization radical ola. ʻO ke kumu ʻelua e hoʻokomo i kahi pūʻulu i loaʻa ka pilina ma waena o ka analyte a me ke kahua analyte ikaika. phenylmaleimide moiety ʻaʻohe uku maʻamau i ka pH maʻamau. Hiki ke hoʻomalu ʻia ka polarity o ka pae paʻa e ka nui o ka styrene a me ka manawa pane o ka polymerization radical manuahi. He koʻikoʻi ka hana hope o ka pane (free-radical polymerization) a hiki ke hoʻololi i ka polarity o ka pae paʻa. ʻO ka hoʻouka ʻana i kēia mau mea paʻa i loko o nā kolamu kila kila a nānā i kā lākou hana chromatographic (selectivity, hoʻonā, N waiwai, a me nā mea ʻē aʻe).
ʻElima mau huila peptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) ua loiloi pū ʻia me ka hoʻohana ʻana i kahi kolamu PMP me ka hoʻohana ʻana i kahi pae kelepona; 60/40 (v/v) acetonitrile/wai ​​(0.1% TFA) ma kahi kahe kahe o 80 μL/min. Ma lalo o nā kūlana elution maikaʻi loa, ʻo ka helu papa helu (N) no kēlā me kēia kolamu (100 × 1.8 mm id) ʻo 20,000 ± 100 (200,000 mau papa TMP / m²). a ua hōʻike ʻia nā chromatograms i ka Figure 5A. ʻO ka loiloi wikiwiki ma kahi kolamu PMP ma ke kahe kahe kiʻekiʻe (700 μL / min), ʻelima mau peptides i hoʻoheheʻe ʻia i loko o hoʻokahi minuke, ua maikaʻi loa nā waiwai N, 13,500 ± 330 i kēlā me kēia kolamu (100 × 1.8 mm id), E like me 0000000000000000000000000000000000000000000000000000000000000000000, ʻO nā kolamu like ʻole (100 × 1.8 mm id) ua hoʻopiha ʻia me ʻekolu mau ʻano like ʻole o ka PMP stationary phase e nānā i ka hana hou ʻana. nā waiwai haʻahaʻa %RSD, e like me ka hōʻike ʻana ma ka Papa 3.
ʻO ka hoʻokaʻawale ʻana o ka hui peptide ma ke kolamu PMP (B) a me ka kolamu Ascentis Express RP-Amide (A); ka pae kelepona 60/40 ACN/H2O (TFA 0.1%), PMP kolamu ana (100 × 1.8 mm id); analytical ʻO ke kauoha elution o nā pūhui: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) a me 5 (leucine) acid enkephalin)).
Ua loiloi ʻia kahi kolamu PMP (100 × 1.8 mm id) no ka hoʻokaʻawale ʻana o nā tryptic digests o ke kanaka serum albumin i ka hana kiʻekiʻe wai chromatography.The chromatogram ma Figure 6 hōʻike ʻia ua hoʻokaʻawale maikaʻi ʻia ka hāpana a maikaʻi loa ka hoʻonā. TFA. E like me ka mea i hōʻike ʻia ma ka chromatogram (Figure 6), ua hoʻokaʻawale ʻia ka HSA digestion i 17 peaks e like me 17 peptides.
Ua hoʻokaʻawale ʻia kahi digest tryptic o HSA (100 × 1.8 mm id) ma kahi kolamu PMP; kahe kahe (100 µL / min), ka pae kelepona 60/40 acetonitrile / wai me 0.1% TFA.
kahi L ka lōʻihi o ke kolamu, η ka viscosity o ka pae kelepona, ΔP ke kolamu hope kaomi, a ʻo u ka linear velocity o ka mobile phase.The permeability o ka PMP kolamu he 2.5 × 10-14 m2, ka kahe kahe ana he 25 μL / min, a me 60/40 v / v / v PMP / meability ua hoʻohana 'ia. mm id) ua like ia me kā mākou noiʻi mua Ref.34. ʻO ka permeability o ke kolamu i hoʻopili ʻia me nā ʻāpana porous superficially: 1.7 × 10-15 no nā ʻāpana 1.3 μm, 3.1 × 10-15 no nā ʻāpana 1.7 μm, 5.2 × 10-15 a me 2.5 μm. μm particles 43. No laila, ua like ka permeability o ka māhele PMP me ka 5 μm core-shell particles.
ʻo Wx ke kaumaha o ke kolamu i hoʻopili ʻia me ka chloroform, ʻo Wy ke kaumaha o ke kolamu i hoʻopili ʻia me ka methanol, a ʻo ρ ka mānoanoa o ka mea hoʻoheheʻe. ʻO nā ʻāpana o ka methanol (ρ = 0.7866) a me ka chloroform (ρ = 1.484). ʻO ka nui o ka porosity o SILICA PARTICLES-C10 × i nā kolamu (103 mm18 ×) ʻO nā kolamu C18-Urea 31 a mākou i aʻo mua ai he 0.63 a me 0.55, ʻo ia hoʻi, ʻo ka hiki ʻana o nā ligands urea e hōʻemi i ka permeability o ka pae paʻa. silica particles no ka mea i loko o ka C18-type stationary phases ua pili ka C18 ligands i na silica particles ma ke ano he mau kaulahao laina, oiai ma ka polystyrene-type stationary phases, ua kukuluia ka A ano mānoanoa polymer layer a puni ia.
Hōʻike ka Figure 7A,B i ke kolamu PMP (100 × 1.8 mm id) a me ka kolamu Ascentis Express RP-Amide (100 × 1.8 mm id) me ka hoʻohana ʻana i nā kūlana elution like (ie, 60/40 ACN/H2O a me 0.1% TFA). ) o ka mokuahi van Deemter. Ua hoʻomākaukau ʻia nā hui peptide i koho ʻia (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) i 20 µL/ ʻO ka liʻiliʻi liʻiliʻi no nā kolamu ʻelua he 800 µL/min. ʻO ke kolamu a me ke kolamu ʻo Ascentis Express RP-Amide he 2.6 µm a me 3.9 µm. ʻO nā helu HETP e hōʻike ana i ka maikaʻi o ka hoʻokaʻawale ʻana o ka kolamu PMP (100 × 1.8 mm id) ʻoi aku ka maikaʻi ma mua o ka mea kūʻai aku ʻo Ascentis Express RP-Amide kolamu (100 i × 1.8). ʻAʻole nui ka emi ʻana o ka waiwai N me ka piʻi ʻana o ke kahe i hoʻohālikelike ʻia i kā mākou noiʻi mua ʻana. ʻO ke kiʻekiʻe o ka hoʻokaʻawale ʻana o ke kolamu PMP (100 × 1.8 mm id) i hoʻohālikelike ʻia i ke kolamu Ascentis Express RP-Amide e pili ana i ka hoʻomaikaʻi ʻana i ke ʻano o ka particle, ka nui, a me nā kaʻina hana hoʻopili kolamu paʻakikī i hoʻohana ʻia i ka hana o kēia manawa34.
(A) van Deemter plot (HETP versus mobile phase linear velocity) i loaʻa me ka PMP column (100 × 1.8 mm id) ma 60/40 ACN/H2O me 0.1% TFA.(B) van Deemter plot (HETP versus mobile phase linear velocity) i loaʻa me ka Ascentis Express RP-Amide velocity 1.00 × 4. ACN/H2O me 0.1% TFA.
Ua hoʻomākaukau a loiloi ʻia kahi papa hana polystyrene i hoʻopaʻa ʻia no ka hoʻokaʻawale ʻana i nā hui peptide synthetic a me nā trypsin digests o ke kanaka serum albumin (HAS) i ka hana kiʻekiʻe wai chromatography. particles, controlled synthesis of the stationary phase, and complex column packing.In addition to high separation efficiency, low column back pressure at high flow rate is another advantage of this stationary phase.PMP columns exhibit good reproducibility and can be used for the analysis of peptide mixtures and trypsin digestion of various proteins.We intend to use this column for the separation of natural bioinal compounds. chromatography.I ka wā e hiki mai ana, e loiloi pū ʻia nā kolamu PMP no ka hoʻokaʻawale ʻana o nā protein a me nā antibodies monoclonal.
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