WO2023129086A2 - Micron-scale biocompatible materials that can bind to negatively charged biomembrane bearing species - Google Patents
Micron-scale biocompatible materials that can bind to negatively charged biomembrane bearing species Download PDFInfo
- Publication number
- WO2023129086A2 WO2023129086A2 PCT/TR2022/051668 TR2022051668W WO2023129086A2 WO 2023129086 A2 WO2023129086 A2 WO 2023129086A2 TR 2022051668 W TR2022051668 W TR 2022051668W WO 2023129086 A2 WO2023129086 A2 WO 2023129086A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- bdpa
- negatively charged
- complexes
- blood
- species
- Prior art date
Links
- 239000000560 biocompatible material Substances 0.000 title description 3
- 239000000463 material Substances 0.000 claims abstract description 51
- 239000008280 blood Substances 0.000 claims abstract description 40
- 210000004369 blood Anatomy 0.000 claims abstract description 40
- 239000007788 liquid Substances 0.000 claims abstract description 36
- 210000004027 cell Anatomy 0.000 claims description 49
- 239000003446 ligand Substances 0.000 claims description 25
- 241000894006 Bacteria Species 0.000 claims description 24
- 241000894007 species Species 0.000 claims description 24
- 239000002245 particle Substances 0.000 claims description 20
- 238000000926 separation method Methods 0.000 claims description 15
- 206010028980 Neoplasm Diseases 0.000 claims description 13
- 201000011510 cancer Diseases 0.000 claims description 13
- 239000011859 microparticle Substances 0.000 claims description 12
- 241000700605 Viruses Species 0.000 claims description 10
- 229920002678 cellulose Polymers 0.000 claims description 9
- 239000001913 cellulose Substances 0.000 claims description 9
- 150000002632 lipids Chemical class 0.000 claims description 9
- 229910052751 metal Inorganic materials 0.000 claims description 9
- 239000002184 metal Substances 0.000 claims description 9
- 230000007704 transition Effects 0.000 claims description 8
- 210000005253 yeast cell Anatomy 0.000 claims description 8
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 claims description 7
- JLJWIVHYUCBSRC-UHFFFAOYSA-N benzene;9-[fluoren-9-ylidene(phenyl)methyl]fluorene Chemical compound C1=CC=CC=C1.C1=CC=CC=C1C([C]1C2=CC=CC=C2C2=CC=CC=C21)=C1C2=CC=CC=C2C2=CC=CC=C21 JLJWIVHYUCBSRC-UHFFFAOYSA-N 0.000 claims description 7
- 150000001768 cations Chemical class 0.000 claims description 7
- 230000007613 environmental effect Effects 0.000 claims description 7
- 235000013305 food Nutrition 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 210000000170 cell membrane Anatomy 0.000 claims description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims description 6
- 150000001412 amines Chemical class 0.000 claims description 4
- 239000003242 anti bacterial agent Substances 0.000 claims description 4
- 229940088710 antibiotic agent Drugs 0.000 claims description 4
- 125000002091 cationic group Chemical group 0.000 claims description 4
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 4
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 4
- LOMZIKCTPMEDDZ-UHFFFAOYSA-N zinc 1-pyridin-2-yl-N-(pyridin-2-ylmethyl)methanamine Chemical compound [Zn+2].C=1C=CC=NC=1CNCC1=CC=CC=N1.C=1C=CC=NC=1CNCC1=CC=CC=N1 LOMZIKCTPMEDDZ-UHFFFAOYSA-N 0.000 claims description 4
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 3
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-O ammonium group Chemical group [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 3
- 229940127557 pharmaceutical product Drugs 0.000 claims description 3
- 239000011574 phosphorus Substances 0.000 claims description 3
- 229910052698 phosphorus Inorganic materials 0.000 claims description 3
- 229920001059 synthetic polymer Polymers 0.000 claims description 3
- RRBYUSWBLVXTQN-UHFFFAOYSA-N tricyclene Chemical compound C12CC3CC2C1(C)C3(C)C RRBYUSWBLVXTQN-UHFFFAOYSA-N 0.000 claims description 3
- RRBYUSWBLVXTQN-VZCHMASFSA-N tricyclene Natural products C([C@@H]12)C3C[C@H]1C2(C)C3(C)C RRBYUSWBLVXTQN-VZCHMASFSA-N 0.000 claims description 3
- 239000002351 wastewater Substances 0.000 claims description 3
- YUFFSWGQGVEMMI-UHFFFAOYSA-N (7Z,10Z,13Z,16Z,19Z)-7,10,13,16,19-docosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCCCC(O)=O YUFFSWGQGVEMMI-UHFFFAOYSA-N 0.000 claims description 2
- YUFFSWGQGVEMMI-JLNKQSITSA-N (7Z,10Z,13Z,16Z,19Z)-docosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCCCC(O)=O YUFFSWGQGVEMMI-JLNKQSITSA-N 0.000 claims description 2
- 239000005864 Sulphur Substances 0.000 claims description 2
- 239000007983 Tris buffer Substances 0.000 claims description 2
- 238000009792 diffusion process Methods 0.000 claims description 2
- XIVFQYWMMJWUCD-UHFFFAOYSA-N dihydrophaseic acid Natural products C1C(O)CC2(C)OCC1(C)C2(O)C=CC(C)=CC(O)=O XIVFQYWMMJWUCD-UHFFFAOYSA-N 0.000 claims description 2
- 239000011521 glass Substances 0.000 claims description 2
- 150000002739 metals Chemical class 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 229920005615 natural polymer Polymers 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims description 2
- 229920000768 polyamine Polymers 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 239000002131 composite material Substances 0.000 claims 1
- 230000001580 bacterial effect Effects 0.000 abstract description 42
- 238000001308 synthesis method Methods 0.000 abstract 1
- 239000002502 liposome Substances 0.000 description 31
- 238000006243 chemical reaction Methods 0.000 description 16
- 239000012528 membrane Substances 0.000 description 15
- 239000004593 Epoxy Substances 0.000 description 13
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 13
- 239000004005 microsphere Substances 0.000 description 12
- 208000031729 Bacteremia Diseases 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 238000001914 filtration Methods 0.000 description 9
- 238000002360 preparation method Methods 0.000 description 9
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 8
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 8
- 239000005090 green fluorescent protein Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 206010040047 Sepsis Diseases 0.000 description 6
- 239000008367 deionised water Substances 0.000 description 6
- 229910021641 deionized water Inorganic materials 0.000 description 6
- 238000002073 fluorescence micrograph Methods 0.000 description 6
- 125000000524 functional group Chemical group 0.000 description 6
- 238000001000 micrograph Methods 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000002105 nanoparticle Substances 0.000 description 4
- 230000007935 neutral effect Effects 0.000 description 4
- 230000003612 virological effect Effects 0.000 description 4
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 3
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- ATBOMIWRCZXYSZ-XZBBILGWSA-N [1-[2,3-dihydroxypropoxy(hydroxy)phosphoryl]oxy-3-hexadecanoyloxypropan-2-yl] (9e,12e)-octadeca-9,12-dienoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCC\C=C\C\C=C\CCCCC ATBOMIWRCZXYSZ-XZBBILGWSA-N 0.000 description 3
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 3
- 239000000356 contaminant Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 241001678559 COVID-19 virus Species 0.000 description 2
- 241000711573 Coronaviridae Species 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 229920002971 Heparan sulfate Polymers 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 238000007605 air drying Methods 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 230000001268 conjugating effect Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- ZRALSGWEFCBTJO-UHFFFAOYSA-O guanidinium Chemical compound NC(N)=[NH2+] ZRALSGWEFCBTJO-UHFFFAOYSA-O 0.000 description 2
- 206010022000 influenza Diseases 0.000 description 2
- GZQKNULLWNGMCW-PWQABINMSA-N lipid A (E. coli) Chemical compound O1[C@H](CO)[C@@H](OP(O)(O)=O)[C@H](OC(=O)C[C@@H](CCCCCCCCCCC)OC(=O)CCCCCCCCCCCCC)[C@@H](NC(=O)C[C@@H](CCCCCCCCCCC)OC(=O)CCCCCCCCCCC)[C@@H]1OC[C@@H]1[C@@H](O)[C@H](OC(=O)C[C@H](O)CCCCCCCCCCC)[C@@H](NC(=O)C[C@H](O)CCCCCCCCCCC)[C@@H](OP(O)(O)=O)O1 GZQKNULLWNGMCW-PWQABINMSA-N 0.000 description 2
- 239000012299 nitrogen atmosphere Substances 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 241000712461 unidentified influenza virus Species 0.000 description 2
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 1
- KXZQYLBVMZGIKC-UHFFFAOYSA-N 1-pyridin-2-yl-n-(pyridin-2-ylmethyl)methanamine Chemical compound C=1C=CC=NC=1CNCC1=CC=CC=N1 KXZQYLBVMZGIKC-UHFFFAOYSA-N 0.000 description 1
- XBNGYFFABRKICK-UHFFFAOYSA-N 2,3,4,5,6-pentafluorophenol Chemical compound OC1=C(F)C(F)=C(F)C(F)=C1F XBNGYFFABRKICK-UHFFFAOYSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 description 1
- BRLQWZUYTZBJKN-UHFFFAOYSA-N Epichlorohydrin Chemical compound ClCC1CO1 BRLQWZUYTZBJKN-UHFFFAOYSA-N 0.000 description 1
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 206010053159 Organ failure Diseases 0.000 description 1
- 206010040070 Septic Shock Diseases 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 239000000538 analytical sample Substances 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000000339 bright-field microscopy Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 229910001431 copper ion Inorganic materials 0.000 description 1
- 239000000412 dendrimer Substances 0.000 description 1
- 229920000736 dendritic polymer Polymers 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 125000003700 epoxy group Chemical group 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000000696 magnetic material Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- IZUPBVBPLAPZRR-UHFFFAOYSA-N pentachloro-phenol Natural products OC1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1Cl IZUPBVBPLAPZRR-UHFFFAOYSA-N 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 238000004094 preconcentration Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 239000004627 regenerated cellulose Substances 0.000 description 1
- 230000036303 septic shock Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- DHCDFWKWKRSZHF-UHFFFAOYSA-N sulfurothioic S-acid Chemical compound OS(O)(=O)=S DHCDFWKWKRSZHF-UHFFFAOYSA-N 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- MVFCZHDZYFNQFW-UHFFFAOYSA-N zinc;1-pyridin-2-yl-n-(pyridin-2-ylmethyl)methanamine Chemical compound [Zn+2].C=1C=CC=NC=1CNCC1=CC=CC=N1 MVFCZHDZYFNQFW-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/022—Filtration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
- A61L2/23—Solid substances, e.g. granules, powders, blocks, tablets
- A61L2/235—Solid substances, e.g. granules, powders, blocks, tablets cellular, porous or foamed
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2202/00—Aspects relating to methods or apparatus for disinfecting or sterilising materials or objects
- A61L2202/10—Apparatus features
- A61L2202/13—Biocide decomposition means, e.g. catalysts, sorbents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2202/00—Aspects relating to methods or apparatus for disinfecting or sterilising materials or objects
- A61L2202/20—Targets to be treated
- A61L2202/22—Blood or products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
- A61M1/3679—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits by absorption
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M2202/00—Special media to be introduced, removed or treated
- A61M2202/20—Pathogenic agents
- A61M2202/203—Bacteria
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
- B01D69/14—Dynamic membranes
- B01D69/141—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes
- B01D69/142—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes with "carriers"
- B01D69/144—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes with "carriers" containing embedded or bound biomolecules
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
- G01N2030/8831—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials involving peptides or proteins
Definitions
- the invention relates to biocompatible micron-scale spherical or non-spherical materials capable of selectively separating bacterial cells from various liquids.
- the invention could potentially be used for the separation and determination of bacterial contamination in pharmaceutical products, food products, and wastewater discharges.
- the invention has application areas such as separating bacteria from the blood of bacteremia patients, separating bacteria from environmental and food samples, and separating species with membrane properties similar to bacteria from similar matrices.
- Bacteremia is the presence of bacteria in the blood, which should normally be sterile. If bacteremia cannot be diagnosed and treated quickly, this may cause sepsis. Sepsis can be defined as the body's immune response to bacterial infection being very severe and the condition damaging healthy tissue. Sepsis may result in septic shock, organ failure, and death. According to CDC data, 1.7 million patients get sepsis in the USA every year, of which approximately 270000 die from sepsis. 30% of patients who die in hospital intensive care units die as a result of sepsis. 15-30% of blood infections worldwide result in death. With the current methods, the clinical diagnosis of bacteremia takes 2-4 days, and its treatment takes 14 days.
- ZnDPA zinc(II)-dipicolylamine
- nanoparticles are derivatized to bind to bacterial contaminants such as Gram-positive bacteria, Gram-negative bacteria, and endotoxins.
- the nanoparticles comprise a core containing a magnetic material and ligands bound to the core.
- Ligands that bind to bacterial contaminants include, for example, a metal ion to form a bis- Zn-DPA or bis-Cu-DPA, e.g., bis(dipicolylamine) (“DPA”) coordinated with Zn 2+ or Cu 2+ .
- Nanoparticles allow the separation of bacterial contaminants when added to liquids such as blood, e.g., whole, or diluted blood, buffer solutions, albumin solutions, beverages for human and/or animal consumption, e.g., drinking water, liquid drugs for humans and/or animals, or other liquids.
- the ZmBDPA complex used in the mentioned invention has a low affinity for negatively charged membranes. It also requires a microfluidic device since nanoparticles are used. Therefore, it is not suitable for applications requiring high flow rates and high volumes, e.g., whole blood filtration.
- the patent document WO 2015/164198 Al provides a method for removing a significant amount of bacteria (e.g., Gram-negative bacteria and Gram-positive bacteria, including bacteria that have no or low affinity for heparan sulfate) from whole blood, serum, or plasma using an adsorption medium.
- bacteria e.g., Gram-negative bacteria and Gram-positive bacteria, including bacteria that have no or low affinity for heparan sulfate
- heparin sulfate can bind some bacterial strains.
- the substrate used it may be a synthetic polymer. It is stated that the substrates used are micron-sized. Whole blood filtration requires micron-scale particles, and this is a known condition.
- the present invention is related to micron-scale spherical and nonspherical materials derivatized with ligands that can selectively bind to bacterial cells, that satisfy the aforementioned requirements, that eliminate all the disadvantages of the prior art and put forward some additional advantages, and that can selectively remove bacterial cells from various liquids, and is also related to the synthesis of said materials.
- the primary object of the invention is the selective separation of bacterial cells from various liquids.
- micron-scale spherical materials derivatized with ligands that can selectively bind to bacterial cells were obtained. It is argued that the micron-scale (50-300 pm) particle size will also allow the flowing liquids to pass through a filtration column consisting of these particles. In this way, bacterial cells can be selectively separated from various liquids, such as blood.
- micron-scale spherical material synthesized by the invention are listed below: i. Suitable for the separation of bacterial cells from high volume liquids. ii. Suitable for large-scale production. iii. Suitable for the separation of bacterial cells from various liquids, including blood. iv. In addition to bacterial cells, they can be used for the separation of circulating tumor cells (i.e., cancer cells circulating in the blood), yeasts, and viral particles (e.g., influenza and SARS-CoV-2) from liquid biological samples, such as blood, and from environmental samples.
- tumor cells i.e., cancer cells circulating in the blood
- yeasts e.g., and viral particles (e.g., influenza and SARS-CoV-2) from liquid biological samples, such as blood, and from environmental samples.
- these materials have been shown to bind to bacteria (1-1.5 micrometers in size) and negatively charged liposomes (150 nm in size). Such materials have the potential to bind to some other species with negatively charged lipid membranes, such as some viruses (e.g., influenza viruses, coronaviruses), some yeast species, and cancer cells circulating in the blood. Therefore, the present invention has many potential applications.
- CM spherical regenerated cellulose microspheres
- ZmBDPA zinc(II) bisdipicolylamine
- phosphate amphiphiles e.g., phosphatidylglycerol, Lipid A, lipoteichoic acid
- the materials obtained by three different chemical approaches are BDPA-CM-I, BDPA-CM-II, and BDPA-CM-III.
- a ligand that can bind to a broad spectrum of bacterial cells is conjugated to the micron-scale particles, and these derivatized particles bind to negatively charged liposomes and bacterial cells.
- Scaled production of spherical and nonspherical microparticles derivatized with ligands capable of selectively binding to bacterial cells is possible and such particles can be used for the removal of bacterial cells from scaled (i.e., large volume) liquids.
- scaled i.e., large volume
- conjugates of micron-scale particles with ZmBDPA complexes were synthesized, and these functional microparticles were shown to bind to negatively charged liposomes and bacterial cells.
- these materials are expected to bind to species bearing negatively charged phosphate lipids similar to bacterial cells such as other microbial pathogens, viruses, and free circulating cancer cells in the blood.
- Products such as cartridges, syringe filters, kits, and other possible devices or apparatus containing the materials synthesized by the invention will be obtained and it will be possible to selectively separate bacteria, viruses, yeast cells, and cancer cells circulating in the blood from various liquids.
- Figure 1 Preparation of CM-epoxy, CM-amine, and BDPA-CM-I.
- Figure 2 Preparation of CM-bisaldehyde and BDPA-CM-II
- Figure 3 Preparation of BDPA-CM-III.
- Figure 5 Bright field micrographs (lOx) for CM-amine (A) and BDPA-CM-I (C) and fluorescence micrographs (lOx, TX-2 filter) for CM-amine (B) and BDPA-CM-I (D) after treatment with negatively charged liposomes and washing.
- Figure 6 Bright field micrographs (lOx) for CM-bisaldehyde (A) and BDPA-CM-II (C) and fluorescence micrographs (lOx, TX-2 filter) for CM-bisaldehyde (B) and BDPA-CM-II (D) after treatment with negatively charged liposomes and washing.
- Figure 7 Bright field micrographs (lOx) for CM-epoxy (A) and BDPA-CM-III (C) and fluorescence micrographs (lOx, TX-2 filter) for CM-epoxy (B) and BDPA-CM-III (D) after treatment with negatively charged liposomes and washing.
- Figure 8 Bright field micrographs (lOx) for CM-amine (A) and BDPA-CM-I (C) and fluorescence micrographs (lOx, GFP filter) for CM-amine (B) and BDPA-CM-I (D) after treatment with GFP expressing Escherichia coli cells and washing.
- Figure 9 Bright field micrographs (lOx) for CM-bisaldehyde (A) and BDPA-CM-II (C) and fluorescence micrographs (lOx, GFP filter) for CM-bisaldehyde (B) and BDPA-CM-II (D) after treatment with GFP expressing Escherichia coli cells and washing.
- Figure 10 Bright field micrographs (lOx) for CM-epoxy (A) and BDPA-CM-III (C) and fluorescence micrographs (lOx, GFP filter) for CM-epoxy (B) and BDPA-CM-III (D) after treatment with GFP expressing Escherichia coli cells and washing.
- biocompatible micron-scale spherical or non-spherical materials capable of selectively separating bacterial cells from various liquids, and their method of preparation and characterization are described only for a better understanding of the subject matter and without any limiting effect.
- the material that can bind to negatively charged biomembrane bearing species is characterized in that it contains microparticles obtained from 20-1000 pm particle-sized materials with a porosity of less than 500 nm, and structures selected from the following groups is bound to the surface of these microparticles: • Zinc(II) bisdipicolylamine (ZmBDPA) complexes or complexes of other transition group metal cations with the BDPA ligand
- the invention relates to biocompatible cellulose microspheres having low porosity (less than 500 nm) decorated with ZmBDPA complexes capable of selectively separating species bearing cell membranes displaying negatively charged lipids from various liquids.
- Species displaying negatively charged lipids on their cell membranes can be liposomes, bacteria, viruses, yeast cells, and cancer cells that circulate in the blood.
- Liquids from which bacteria and species with similar membrane properties can be separated include blood, plasma, food, environmental samples, and pharmaceutical products and formulations.
- species bearing cell membranes displaying negatively charged lipids can be separated from liquids by using microspheres manufactured from other materials with low porosity, such as glass, synthetic and natural polymers.
- the species can be separated from liquids which such materials can be liposomes, bacteria, viruses, yeast cells, and cancer cells that circulate in the blood.
- the object of the invention is to separate bacterial cells from liquids using micron-scale spherical materials derivatized with ligands (ZmBDPA complexes) that can selectively bind to bacterial cells (and species with membrane structures carrying similar negatively charged phosphate amphiphiles).
- the ligands bound to the particles provide selective binding.
- the micron-scale (50-300 pm) particle size allows the flowing liquids to pass through a filtration column of these particles. In this way, bacterial cells can be selectively separated from various liquids, such as blood.
- Such materials have the potential to bind to some other species with negatively charged lipid membranes, such as some viruses (e.g., influenza viruses, coronaviruses), some yeast species, and cancer cells circulating in the blood.
- Zn 2+ zinc ion
- Cu 2+ copper ion
- the microsphere-bound recognition group i.e., the ZmBDPA ligand in the prototype material
- CM regenerated spherical cellulose microspheres
- ZmBDPA zinc(II) bisdipicolylamine
- BDPA-CM-I was obtained by conjugation of BDPA-Acid and CM- Amine ( Figure 1).
- BDPA-CM-II was obtained from the reaction of CM-Bisaldehyde with BDPA- Amine ( Figure 2).
- BDPA-CM-III was obtained from the reaction of CM-Epoxy with BDPA-Amine ( Figure 3). These materials are characterized by FTIR and elemental analysis.
- CM-Epoxy 0.5 mL of epichlorohydrin and 1.5 mL of 2.5 M aqueous NaOH solution were added to 500 mg cellulose microspheres. The reaction was allowed to proceed for 2.5 hours at 40°C in a shaking incubator at 250 rpm. Following the reaction, the microspheres were filtered and washed with deionized water and acetone. After air drying for 24 hours, CM- epoxy was lyophilized. The oxirane concentration of CM-epoxy was determined by the thiosulfate titration method. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1580, 1237, 1022, 896, 796.7 cm' 1 .
- CM-Amine 1 mL of 25% NFL OH solution was added to 100 mg CM-epoxy in the test tube and the reaction was continued at room temperature for 16 hours in a shaking incubator at 150 rpm. Following the reaction, the microspheres were filtered and washed with deionized water and acetone. After air drying for 24 hours, CM-amine was lyophilized. The lyophilized CM- amine spheres were examined by CHNS analysis and the amine group concentration in the spheres was in the range of 257-421 pmol/g. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1427, 1050 cm' 1 .
- BDPA-CM-I To 25 mg BDPA-acid in 2.5 mL DMF (dimethylformamide) was added 7.19 mg (0.039 mmol) of PfpOH (pentafluorophenol), 7.47 mg (0.039 mmol) of EDC (l-ethyl-3- (3-dimethylaminopropyl)carbodiimide), and 1 mg (0.007 mmol) of DMAP (p-N,N- dimethylaminopyridine) at 0°C, respectively. After the reaction was continued at 0°C for 10 minutes, it was brought to room temperature and continued at this temperature for 16 hours.
- PfpOH pentethyl-3- (3-dimethylaminopropyl)carbodiimide
- DMAP p-N,N- dimethylaminopyridine
- CM-Bisaldehyde 1.5 mL 0.20 M NalOi (in water) was added to 488.1 mg cellulose microspheres, and the reaction was continued for 24 hours at room temperature in a shaking incubator at 250 rpm in the dark. After the reaction, the microspheres were filtered and washed five times with deionized water. CM-Bisaldehyde spheres were stored at 4°C until use. An analytical sample was lyophilized and the content of aldehyde by titration was found in the range of 5.79-12.87 mmol/g. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1681, 894 cm' 1 .
- BDPA-CM-II 100 mg (0.2 mmol) of BDPA-amine in 5 mL MeOH (methanol) was added to 100 mg (1.3 mmol aldehyde functional group) CM-bisaldehyde and the reaction was continued for 16 hours at room temperature in a shaking incubator at 250 rpm. 38.6 mg (1 mmol) of NaBEL (sodium borohydride) was added to the reaction and the reaction was continued with 250 rpm agitation at room temperature for 4 hours. After filtration, the spheres were washed with 1x5 mL of 1 M NaOH and 3x25 mL of deionized water, and 1x25 mL of ethanol.
- NaBEL sodium borohydride
- the BDPA-CM-II obtained was lyophilized and the BDPA ligand concentration in the spheres was found to be 0.070 pmol/mg by CHNS analysis. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1592, 1156, 1026, 894, 798 cm' 1 .
- BDPA-CM-III 50 mg (83.8 pmol) BDPA-amine was added to 100 mg (83.8 pmol epoxy group containing) CM-Epoxy in 1 mL of acetic acid and the reaction was boiled using a reflux condenser under nitrogen atmosphere for 16 hours. The reaction was washed with 20 mL of ethanol, 10 mL of 0.1 M HC1, 10 mL of 0.1 M NaOH, and 10 mL of deionized water. The BDPA-CM-III obtained was lyophilized and the BDPA ligand concentration in the spheres was found to be 0.020 pmol/mg by CHNS analysis. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1596, 1157, 1023, 796 cm' 1 .
- phosphatidylcholine (68%), cholesterol (30%), and DiO fluorescent dye (2%)
- negatively charged liposomes were made from phosphatidylglycerol (10%) which is a negatively charged lipid, phosphatidylcholine (58%), cholesterol (30%), and DiD fluorescent (red) dye (2%).
- the diameters of the liposomes used are between 140-180 nm on average.
- BDPA-CM-II binds to a slightly lesser amount of negatively charged liposomes.
- GFP green fluorescent protein
- such materials can be used to separate cancer cells circulating in the blood, yeasts, and viral particles (e.g., influenza and SARS-CoV-2), all of which have negatively charged cell membranes, from liquid biological and environmental samples.
- viral particles e.g., influenza and SARS-CoV-2
- cellulose which has been used in the preparation of the prototype material
- other polymeric materials can be used as the support material.
- complexes of other transition group metal cations with BDPA or ligands with other structures, cationic groups, peptides, antibiotics, and antibodies can be used as affinity ligands.
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to biocompatible micron-scale spherical or non-spherical materials that can selectively separate bacterial cells from blood and various other liquids, and the synthesis methods thereof.
Description
MICRON-SCALE BIOCOMPATIBLE MATERIALS THAT CAN BIND TO
NEGATIVELY CHARGED BIOMEMBRANE BEARING SPECIES
Technical Field of the Invention
The invention relates to biocompatible micron-scale spherical or non-spherical materials capable of selectively separating bacterial cells from various liquids. The invention could potentially be used for the separation and determination of bacterial contamination in pharmaceutical products, food products, and wastewater discharges. The invention has application areas such as separating bacteria from the blood of bacteremia patients, separating bacteria from environmental and food samples, and separating species with membrane properties similar to bacteria from similar matrices.
State of the Art of the Invention (Prior Art)
Bacteremia is the presence of bacteria in the blood, which should normally be sterile. If bacteremia cannot be diagnosed and treated quickly, this may cause sepsis. Sepsis can be defined as the body's immune response to bacterial infection being very severe and the condition damaging healthy tissue. Sepsis may result in septic shock, organ failure, and death. According to CDC data, 1.7 million patients get sepsis in the USA every year, of which approximately 270000 die from sepsis. 30% of patients who die in hospital intensive care units die as a result of sepsis. 15-30% of blood infections worldwide result in death. With the current methods, the clinical diagnosis of bacteremia takes 2-4 days, and its treatment takes 14 days.
In the state of the art, there are materials developed for the separation of bacterial cells in the blood.
Bradley D. Smith et al. described the development of synthetic zinc(II)-dipicolylamine (ZnDPA) receptors as selective targeting agents for anionic membranes in cell culture and living experimental subjects. ZmBDPA systems have been known to target bacterial
membranes since at least 2006. The preparation of micron-scale support materials carrying ZmBDPA complexes and the separation of negatively charged membrane-carrying species (liposomes, bacterial cells, viruses, yeast cells, cancer cells) from liquids are not mentioned in this document.
In patent document US 2014/0212335 Al, nanoparticles are derivatized to bind to bacterial contaminants such as Gram-positive bacteria, Gram-negative bacteria, and endotoxins. The nanoparticles comprise a core containing a magnetic material and ligands bound to the core. Ligands that bind to bacterial contaminants include, for example, a metal ion to form a bis- Zn-DPA or bis-Cu-DPA, e.g., bis(dipicolylamine) (“DPA”) coordinated with Zn2+ or Cu2+. Nanoparticles allow the separation of bacterial contaminants when added to liquids such as blood, e.g., whole, or diluted blood, buffer solutions, albumin solutions, beverages for human and/or animal consumption, e.g., drinking water, liquid drugs for humans and/or animals, or other liquids. The ZmBDPA complex used in the mentioned invention has a low affinity for negatively charged membranes. It also requires a microfluidic device since nanoparticles are used. Therefore, it is not suitable for applications requiring high flow rates and high volumes, e.g., whole blood filtration. Some studies have shown that dendrimers, dyes, and liposomes carrying ZmBDPA can selectively bind to bacterial cells in vivo and in vitro [1-4],
The patent document WO 2015/164198 Al provides a method for removing a significant amount of bacteria (e.g., Gram-negative bacteria and Gram-positive bacteria, including bacteria that have no or low affinity for heparan sulfate) from whole blood, serum, or plasma using an adsorption medium. The main invention in this document is that heparin sulfate can bind some bacterial strains. Although it is not clear what the substrate used is, it may be a synthetic polymer. It is stated that the substrates used are micron-sized. Whole blood filtration requires micron-scale particles, and this is a known condition.
None of the known systems in the state of the art is suitable for the separation of negatively charged membrane carrying species (liposomes, bacterial cells, yeast cells, free circulating cancer cells, and some virus particles) from liquids of various volumes (from milliliters to liters) flowing at high flow rates and for complete (i.e., high volume) blood filtration.
Brief Description and Objects of the Invention
The present invention is related to micron-scale spherical and nonspherical materials derivatized with ligands that can selectively bind to bacterial cells, that satisfy the aforementioned requirements, that eliminate all the disadvantages of the prior art and put forward some additional advantages, and that can selectively remove bacterial cells from various liquids, and is also related to the synthesis of said materials.
With this invention, it is possible to separate negatively charged membrane-carrying species (liposomes, bacterial cells, yeast cells, free circulating cancer cells, and some virus particles) from liquids (blood, diluted blood, food samples, drug samples, waste waters, etc.) of various volumes (from milliliters to liters) flowing at any flow rate above the diffusion limit or a high flow rate of 0.0001 mL/min to 100000 mL/min by derivatizing micron-scale, spherical or non-spherical and biocompatible materials with ZmBDPA ligands. The same separation process with the materials of the invention can also be performed with still liquids.
The primary object of the invention is the selective separation of bacterial cells from various liquids. For this purpose, micron-scale spherical materials derivatized with ligands that can selectively bind to bacterial cells were obtained. It is argued that the micron-scale (50-300 pm) particle size will also allow the flowing liquids to pass through a filtration column consisting of these particles. In this way, bacterial cells can be selectively separated from various liquids, such as blood.
The advantages of the micron-scale spherical material synthesized by the invention are listed below: i. Suitable for the separation of bacterial cells from high volume liquids. ii. Suitable for large-scale production. iii. Suitable for the separation of bacterial cells from various liquids, including blood. iv. In addition to bacterial cells, they can be used for the separation of circulating tumor cells (i.e., cancer cells circulating in the blood), yeasts, and viral particles (e.g., influenza and
SARS-CoV-2) from liquid biological samples, such as blood, and from environmental samples.
With this invention, it is possible to selectively separate bacteria and species with similar membrane properties from liquids. By immobilizing ZmBDPA complexes, which have the capability of selectively binding to bacteria (and other species with similar membrane properties), onto low porosity microparticles (in this case cellulose microparticles) it is possible to separate said species from liquids flowing at a certain rate. In this way, by using whole blood filtration, the concentration of bacteria in the blood of bacteremia patients can be reduced and the patients can be cured. In the same way, bacteria in analytical blood samples can be separated using such particles and bacteremia diagnosis can be facilitated. The materials of the invention can also be used both for the large-scale separation of bacteria from similarly flowing environmental and food samples and for the separation of bacteria from analytical-scale samples. During the invention studies, these materials have been shown to bind to bacteria (1-1.5 micrometers in size) and negatively charged liposomes (150 nm in size). Such materials have the potential to bind to some other species with negatively charged lipid membranes, such as some viruses (e.g., influenza viruses, coronaviruses), some yeast species, and cancer cells circulating in the blood. Therefore, the present invention has many potential applications.
Prototype materials developed for this purpose were obtained by appropriately derivatizing spherical regenerated cellulose microspheres (CM, diameter 50-75 pm) known to be biocompatible and conjugating them with zinc(II) bisdipicolylamine (ZmBDPA) complexes that are known to bind selectively to negatively charged phosphate amphiphiles (e.g., phosphatidylglycerol, Lipid A, lipoteichoic acid) on the surfaces of bacterial cells. The materials obtained by three different chemical approaches are BDPA-CM-I, BDPA-CM-II, and BDPA-CM-III.
With the invention, a ligand that can bind to a broad spectrum of bacterial cells is conjugated to the micron-scale particles, and these derivatized particles bind to negatively charged liposomes and bacterial cells. Scaled production of spherical and nonspherical microparticles derivatized with ligands capable of selectively binding to bacterial cells is possible and such particles can be used for the removal of bacterial cells from scaled (i.e., large volume) liquids.
In addition, it will be possible to diagnose bacteremia rapidly with cartridges, columns, or kits containing these materials and to perform bacteremia treatment with full blood filtration. With the resulting prototype material, in addition to bacteria, it will be possible for cancer cells, some yeast cells, and some viral particles circulating in the blood to be separated from blood and other biological liquids. With these particles, it is possible to separate the relevant species not only from still liquids but also from flowing liquids.
With the invention, conjugates of micron-scale particles with ZmBDPA complexes were synthesized, and these functional microparticles were shown to bind to negatively charged liposomes and bacterial cells. In addition, these materials are expected to bind to species bearing negatively charged phosphate lipids similar to bacterial cells such as other microbial pathogens, viruses, and free circulating cancer cells in the blood.
It will be possible to quickly determine blood infections and to quickly separate the bacteria in blood with the materials of the invention. In addition, a large number of applications requiring bacterial separation will be possible with these materials. Species such as yeast and viral particles with lipid membranes similar to bacteria can also be separated from biological and other liquids with these materials.
Products such as cartridges, syringe filters, kits, and other possible devices or apparatus containing the materials synthesized by the invention will be obtained and it will be possible to selectively separate bacteria, viruses, yeast cells, and cancer cells circulating in the blood from various liquids.
Definitions of Figures Describing the Invention
The figures and related descriptions required to better understand the subject of the invention are as follows.
Figure 1: Preparation of CM-epoxy, CM-amine, and BDPA-CM-I.
Figure 2: Preparation of CM-bisaldehyde and BDPA-CM-II Figure 3: Preparation of BDPA-CM-III.
Figure 4: Normalized fluorescence data after incubation of liposomal dispersions with CMs (Liposome A = neutral liposomes, Liposome B = negatively charged liposomes).
Figure 5: Bright field micrographs (lOx) for CM-amine (A) and BDPA-CM-I (C) and fluorescence micrographs (lOx, TX-2 filter) for CM-amine (B) and BDPA-CM-I (D) after treatment with negatively charged liposomes and washing.
Figure 6: Bright field micrographs (lOx) for CM-bisaldehyde (A) and BDPA-CM-II (C) and fluorescence micrographs (lOx, TX-2 filter) for CM-bisaldehyde (B) and BDPA-CM-II (D) after treatment with negatively charged liposomes and washing.
Figure 7: Bright field micrographs (lOx) for CM-epoxy (A) and BDPA-CM-III (C) and fluorescence micrographs (lOx, TX-2 filter) for CM-epoxy (B) and BDPA-CM-III (D) after treatment with negatively charged liposomes and washing.
Figure 8: Bright field micrographs (lOx) for CM-amine (A) and BDPA-CM-I (C) and fluorescence micrographs (lOx, GFP filter) for CM-amine (B) and BDPA-CM-I (D) after treatment with GFP expressing Escherichia coli cells and washing.
Figure 9: Bright field micrographs (lOx) for CM-bisaldehyde (A) and BDPA-CM-II (C) and fluorescence micrographs (lOx, GFP filter) for CM-bisaldehyde (B) and BDPA-CM-II (D) after treatment with GFP expressing Escherichia coli cells and washing.
Figure 10: Bright field micrographs (lOx) for CM-epoxy (A) and BDPA-CM-III (C) and fluorescence micrographs (lOx, GFP filter) for CM-epoxy (B) and BDPA-CM-III (D) after treatment with GFP expressing Escherichia coli cells and washing.
Detailed Description of the Invention
In this detailed description, biocompatible micron-scale spherical or non-spherical materials capable of selectively separating bacterial cells from various liquids, and their method of preparation and characterization are described only for a better understanding of the subject matter and without any limiting effect.
The material that can bind to negatively charged biomembrane bearing species, is characterized in that it contains microparticles obtained from 20-1000 pm particle-sized materials with a porosity of less than 500 nm, and structures selected from the following groups is bound to the surface of these microparticles:
• Zinc(II) bisdipicolylamine (ZmBDPA) complexes or complexes of other transition group metal cations with the BDPA ligand
• Complexes of zinc (II) and other transition group metal cations with DPA, cyclene, or imidazole (bis, tris, tetra, or poly) ligands
• Complexes obtained with ligand structures containing nitrogen, phosphorus, sulphur or oxygen, or mixtures thereof, known to form complexes with zinc (II) and other transition group metals
• Cationic groups of amine, polyamine, ammonium, polyammonium, guanidinium, or polyguanidinium,
• Peptides, proteins, antibiotics, or antibodies.
The invention relates to biocompatible cellulose microspheres having low porosity (less than 500 nm) decorated with ZmBDPA complexes capable of selectively separating species bearing cell membranes displaying negatively charged lipids from various liquids. Species displaying negatively charged lipids on their cell membranes can be liposomes, bacteria, viruses, yeast cells, and cancer cells that circulate in the blood. Liquids from which bacteria and species with similar membrane properties can be separated include blood, plasma, food, environmental samples, and pharmaceutical products and formulations.
Apart from cellulose decorated with ZmBDPA complexes, species bearing cell membranes displaying negatively charged lipids can be separated from liquids by using microspheres manufactured from other materials with low porosity, such as glass, synthetic and natural polymers. The species can be separated from liquids which such materials can be liposomes, bacteria, viruses, yeast cells, and cancer cells that circulate in the blood.
The object of the invention is to separate bacterial cells from liquids using micron-scale spherical materials derivatized with ligands (ZmBDPA complexes) that can selectively bind to bacterial cells (and species with membrane structures carrying similar negatively charged phosphate amphiphiles). The ligands bound to the particles provide selective binding. The micron-scale (50-300 pm) particle size allows the flowing liquids to pass through a filtration column of these particles. In this way, bacterial cells can be selectively separated from various liquids, such as blood. Such materials have the potential to bind to some other species with
negatively charged lipid membranes, such as some viruses (e.g., influenza viruses, coronaviruses), some yeast species, and cancer cells circulating in the blood.
In the structure of the ZmBDPA complex ligand, instead of Zn2+ (zinc ion) as the metal cation, there may be Cu2+ (copper ion) or other transition metals with stable oxidation numbers of +2 and +3.
In the ligand structure, instead of BDPA, there may be ligand structures containing DPA, cyclene, bis- or tetraimidazole, or nitrogen, phosphorus, sulfur, or oxygen that can form complexes with transition elements with stable oxidation numbers of +2 and +3.
The microsphere-bound recognition group (i.e., the ZmBDPA ligand in the prototype material) may additionally be cationic groups (amine, ammonium, guanidinium, etc.), antibodies, peptides, antibiotics (e.g., vancomycin), or carbohydrates that can bind to bacterial cells and species with similar membrane structures.
Prototype materials developed for this purpose were obtained by appropriately derivatizing regenerated spherical cellulose microspheres (CM, diameter 50-75 pm) known to be biocompatible and conjugating them with zinc(II) bisdipicolylamine (ZmBDPA) complexes known to bind selectively to negatively charged phosphate amphiphiles (e.g., phosphatidylglycerol, Lipid A, lipoteichoic acid) on the surfaces of bacterial cells. The materials obtained by three different chemical approaches are BDPA-CM-I, BDPA-CM-II, and BDPA-CM-III. BDPA-CM-I was obtained by conjugation of BDPA-Acid and CM- Amine (Figure 1). BDPA-CM-II was obtained from the reaction of CM-Bisaldehyde with BDPA- Amine (Figure 2). BDPA-CM-III was obtained from the reaction of CM-Epoxy with BDPA-Amine (Figure 3). These materials are characterized by FTIR and elemental analysis.
Procedures and Characterization Data for BDPA-CM-I, BDPA-CM-II, and BDPA-CM- III
CM-Epoxy: 0.5 mL of epichlorohydrin and 1.5 mL of 2.5 M aqueous NaOH solution were added to 500 mg cellulose microspheres. The reaction was allowed to proceed for 2.5 hours at 40°C in a shaking incubator at 250 rpm. Following the reaction, the microspheres were filtered and washed with deionized water and acetone. After air drying for 24 hours, CM- epoxy was lyophilized. The oxirane concentration of CM-epoxy was determined by the thiosulfate titration method. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1580, 1237, 1022, 896, 796.7 cm'1.
CM-Amine: 1 mL of 25% NFL OH solution was added to 100 mg CM-epoxy in the test tube and the reaction was continued at room temperature for 16 hours in a shaking incubator at 150 rpm. Following the reaction, the microspheres were filtered and washed with deionized water and acetone. After air drying for 24 hours, CM-amine was lyophilized. The lyophilized CM- amine spheres were examined by CHNS analysis and the amine group concentration in the spheres was in the range of 257-421 pmol/g. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1427, 1050 cm'1.
BDPA-CM-I: To 25 mg BDPA-acid in 2.5 mL DMF (dimethylformamide) was added 7.19 mg (0.039 mmol) of PfpOH (pentafluorophenol), 7.47 mg (0.039 mmol) of EDC (l-ethyl-3- (3-dimethylaminopropyl)carbodiimide), and 1 mg (0.007 mmol) of DMAP (p-N,N- dimethylaminopyridine) at 0°C, respectively. After the reaction was continued at 0°C for 10
minutes, it was brought to room temperature and continued at this temperature for 16 hours. 5.05 mg (6.81 pL, 0.039 mmol) of DIPEA (ethyldiisopropylamine) and 35 mg (containing 343 pmol/g of amine) of CM-amine were added to the reaction in 0.5 mL DMF, respectively. The reaction was continued for 16 hours at room temperature and under a nitrogen atmosphere. After the solvent was removed under low pressure, BDPA-CM-I spheres were washed with 3x25 mL deionized water and 1x25 mL ethanol. Lyophilized BDPA-CM-I spheres were examined by CHNS analysis and the BDPA ligand concentration in the spheres was found to be 0.123 pmol/mg. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1651, 1061, 894, 766 cm'1.
CM-Bisaldehyde: 1.5 mL 0.20 M NalOi (in water) was added to 488.1 mg cellulose microspheres, and the reaction was continued for 24 hours at room temperature in a shaking incubator at 250 rpm in the dark. After the reaction, the microspheres were filtered and washed five times with deionized water. CM-Bisaldehyde spheres were stored at 4°C until use. An analytical sample was lyophilized and the content of aldehyde by titration was found in the range of 5.79-12.87 mmol/g. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1681, 894 cm'1.
BDPA-CM-II: 100 mg (0.2 mmol) of BDPA-amine in 5 mL MeOH (methanol) was added to 100 mg (1.3 mmol aldehyde functional group) CM-bisaldehyde and the reaction was continued for 16 hours at room temperature in a shaking incubator at 250 rpm. 38.6 mg (1 mmol) of NaBEL (sodium borohydride) was added to the reaction and the reaction was continued with 250 rpm agitation at room temperature for 4 hours. After filtration, the spheres were washed with 1x5 mL of 1 M NaOH and 3x25 mL of deionized water, and 1x25 mL of ethanol. The BDPA-CM-II obtained was lyophilized and the BDPA ligand concentration in the spheres was found to be 0.070 pmol/mg by CHNS analysis. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1592, 1156, 1026, 894, 798 cm'1.
CM-Epoxy BDPA-Amine BDPA-CM-III
BDPA-CM-III: 50 mg (83.8 pmol) BDPA-amine was added to 100 mg (83.8 pmol epoxy group containing) CM-Epoxy in 1 mL of acetic acid and the reaction was boiled using a reflux condenser under nitrogen atmosphere for 16 hours. The reaction was washed with 20 mL of ethanol, 10 mL of 0.1 M HC1, 10 mL of 0.1 M NaOH, and 10 mL of deionized water. The BDPA-CM-III obtained was lyophilized and the BDPA ligand concentration in the spheres was found to be 0.020 pmol/mg by CHNS analysis. Functional groups in the spheres were determined by FTIR. FTIR (KBr) v 1596, 1157, 1023, 796 cm'1.
Two different studies were conducted to determine the affinity of BDPA-CM-I, BDPA-CM- II, and BDPA-CM-III to negatively charged membranes. In the first study, the binding of these conjugates and controls (CM-Amine for BDPA-CM-I, CM-Bisaldehyde for BDPA-CM- II, CM-Epoxy for BDPA-CM-III) to negatively charged liposomes was examined. In liposome studies, neutral liposomes were made from phosphatidylcholine (68%), cholesterol
(30%), and DiO fluorescent dye (2%), and negatively charged liposomes were made from phosphatidylglycerol (10%) which is a negatively charged lipid, phosphatidylcholine (58%), cholesterol (30%), and DiD fluorescent (red) dye (2%). The diameters of the liposomes used are between 140-180 nm on average. It was observed that BDPA-CMs and their respective controls did not significantly bind to neutral liposomes, negatively charged liposomes were bound in significant amounts by BDPA-CM-I, BDPA-CM-II, and BDPA-CM-III, but not in significant amounts by their associated controls (Figure 4, Liposome A = neutral liposomes, Liposome B = negatively charged liposomes). Fluorescence microscopy has also shown that BDPA-CM-I, BDPA-CM-II, and BDPA-CM-III bind to negatively charged liposomes (Figures 5, 6, and 7). It can be calculated that BDPA-CM-I and BDPA-CM-III particles with an average diameter of 62 pm each bind an average of 69000 liposomes per microsphere. BDPA-CM-II binds to a slightly lesser amount of negatively charged liposomes. In the second study conducted to determine the affinity of BDPA-CM-I, BDPA-CM-II, and BDPA-CM-III to negatively charged membranes, the binding of these conjugates and their controls to GFP (green fluorescent protein) expressing Escherichia coli bacterial cells was examined. CMs were incubated with bacterial dispersions, washed, and examined with bright field and fluorescence microscopy. In this study, it was observed that BDPA-CMs were able to bind to bacterial cells much better than their controls (Figure 8, 9, 10).
These studies have shown that cellulose microspheres with diameters of 50-75 pm, derivatized with a ligand that can bind to bacterial cells, can bind to negatively charged liposomes and bacterial cells. Some potential application areas of the materials subject to this this invention are listed below:
• Preconcentration of bacterial cells by separating them from blood for rapid diagnosis of bacteremia that may be defined as bacteria in the blood or blood infection
• Whole blood filtration with columns containing these materials for the treatment of bacteremia
• Separation of bacterial cells from environmental, food, and drug samples.
In addition to these potential applications, such materials can be used to separate cancer cells circulating in the blood, yeasts, and viral particles (e.g., influenza and SARS-CoV-2), all of
which have negatively charged cell membranes, from liquid biological and environmental samples.
In addition to cellulose, which has been used in the preparation of the prototype material, other polymeric materials can be used as the support material. In addition to the ZmBDPA complexes used in the prototype material, complexes of other transition group metal cations with BDPA or ligands with other structures, cationic groups, peptides, antibiotics, and antibodies can be used as affinity ligands.
References
1. Rice, D.; Plaunt, A. J.; Turkyilmaz, S.; Smith, M.; Wang, Y.; Rusckowski, M.; Smith B.D. Evaluation of [11 Iln]-Labeled Zinc-Dipicolylamine Tracers for SPECT Imaging of Bacterial Infection. Mol. Imaging Biol. 2015, 77, 204-213
2. Turkyilmaz, S.; Rice, D.R.; Palumbo, R.; Smith B.D. Selective recognition of anionic cell membranes using targeted liposomes coated with zinc(II)-bis(dipicolylamine) affinity units. Org. Biomol. Chem. 2014, 12, 5645-5655
3. Xiao, S.; Abu-Esba, L.; Turkyilmaz, S.; White, A.G.; Smith B.D. Multivalent Dendritic Molecules as Broad Spectrum Bacteria Agglutination Agents. Theranostics 2013, 3 (9), 658-666
4. Xiao, S.; Turkyilmaz, S.; Smith, B.D. Convenient Synthesis of Multivalent Zinc(II)- Dipicolylamine Complexes as Potential Cell Targeting Agents. Tetrahedron Let. 2013, 54, 861-864
Claims
1. A material that can bind to negatively charged biomembrane bearing species, comprising microparticles obtained from a material with a porosity of less than 500 nm and are in 20-1000 pm particle size range, wherein the structure selected from the following group is bound to the surface of these microparticles:
• Zinc(II) bisdipicolylamine (ZmBDPA) complexes or complexes of other transition group metal cations with BDPA ligands
• Complexes of zinc (II) and other transition group metal cations with DPA, cyclene, or imidazole (bis, tris, tetra, or poly) ligands
• Complexes obtained with ligand structures containing nitrogen, phosphorus, sulphur or oxygen or mixtures thereof known to form complexes with zinc (II) and other transition group metal cations
• Cationic groups like amines, polyamines, ammoniums, polyammoniums, guanidiniums, or polyguanidiniums,
• Peptides, proteins, antibiotics, or antibodies.
2. A material according to claim 1, wherein the material has a spherical or non-spherical shape.
3. A material according to claim 1, wherein the microparticles are microparticles obtained from cellulose.
4. A material according to claim 1, wherein the microparticles are microparticles obtained from glass, synthetic and natural polymers, metals, or composites of these materials.
5. A material according to claim 1, which are used in the separation of the species with cell membranes displaying negatively charged lipids from liquids with any flow rate above the diffusion limit or 0.0001 mL/min and under 100000 mL/min or from still liquids.
6. A material according to claim 5, wherein the liquid is blood, plasma, food, environmental samples, wastewater discharges, pharmaceutical products, or pharmaceutical formulations.
7. A material according to claim 1, wherein the species with cell membranes displaying negatively charged lipids are bacteria, viruses, yeast cells or cancer cells circulating in the blood.
8. An apparatus comprising the material of any one of the preceding claims.
9. A device comprising the material of any one of the preceding claims.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
TR2021021980 | 2021-12-30 | ||
TR2021/021980 TR2021021980A1 (en) | 2021-12-30 | MICRON SCALE BIOCOMPATIBLE MATERIALS THAT CAN RETAIN NEGATIVELY CHARGEED BIOMEMEMBRANE-BEARING SPECIES |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2023129086A2 true WO2023129086A2 (en) | 2023-07-06 |
WO2023129086A3 WO2023129086A3 (en) | 2024-04-18 |
Family
ID=87000436
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/TR2022/051668 WO2023129086A2 (en) | 2021-12-30 | 2022-12-29 | Micron-scale biocompatible materials that can bind to negatively charged biomembrane bearing species |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO2023129086A2 (en) |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9566355B2 (en) * | 2013-03-13 | 2017-02-14 | Northwestern University | Bacteria-targeted magnetic resonance contrast agents |
-
2022
- 2022-12-29 WO PCT/TR2022/051668 patent/WO2023129086A2/en unknown
Also Published As
Publication number | Publication date |
---|---|
WO2023129086A3 (en) | 2024-04-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Cai et al. | Immuno-modified superparamagnetic nanoparticles via host–guest interactions for high-purity capture and mild release of exosomes | |
US11771811B2 (en) | Method and device for purification of blood from circulating cell free DNA | |
DE60306135T2 (en) | IMPROVED SEPARATION | |
Pranantyo et al. | Antimicrobial peptide-reduced gold nanoclusters with charge-reversal moieties for bacterial targeting and imaging | |
EP2389242B1 (en) | Sorbent for endotoxins | |
Rabiee et al. | Aptamer hybrid nanocomplexes as targeting components for antibiotic/gene delivery systems and diagnostics: a review | |
WO2006012884A1 (en) | Device and method for isolating cells, bioparticles and/or molecules from liquids designed to be used for the treatment of human diseases | |
CN106457205A (en) | Adsorbent for removing histone and purification device for liquid derived from living organism | |
SE526038C2 (en) | Polymer affinity matrix, process for its preparation and use thereof | |
JP2022141661A (en) | Aqueous biomolecule coupling on co2-plasma-activated surfaces | |
US20100190690A1 (en) | Biomimetic particles and films for pathogen capture and other uses | |
DE102005036505A1 (en) | Filtration process and assembly for removing harmful substances from human blood e.g. human immunodeficiency virus, Herpes virus, or plasmodia, comprises a reactor vessel of a biocompatible material | |
Chen et al. | Assembled hemoglobin and catalase nanotubes for the treatment of oxidative stress | |
Wang et al. | Cytotoxicity evaluation of pH-controlled antitumor drug release system of titanium dioxide nanotubes | |
Pan et al. | Rapid and efficient isolation platform for plasma extracellular vesicles: EV‐FISHER | |
DE102004054536A1 (en) | Multimodal modified cells as cellular dosage forms for active substances and as diagnostic cell particles | |
WO2023129086A2 (en) | Micron-scale biocompatible materials that can bind to negatively charged biomembrane bearing species | |
CN103301446B (en) | Affinity adsorption material for treating hyperbilirubinemia and preparation method thereof | |
TR2021021980A1 (en) | MICRON SCALE BIOCOMPATIBLE MATERIALS THAT CAN RETAIN NEGATIVELY CHARGEED BIOMEMEMBRANE-BEARING SPECIES | |
DE102005063175A1 (en) | Separation of cells and bio-particles and/or molecules, for animals and biotechnology and medical diagnosis, adds functionalized micro-particles for recognition and bonding to be separated by membranes | |
CN115554314B (en) | Protein manganese composition for resisting liver fibrosis | |
CN108371714A (en) | A kind of stannic oxide/graphene nano passs medicine process and its application | |
JP2006333850A (en) | Base material for separating biological particle | |
Razdan | Polymeric nanoparticle as a new platform for high removal of endotoxins from biopharmaceutical solutions | |
Kuypers et al. | A functional assessment of affinity isolated inflammatory extracellular vesicle subpopulations |