Claims (12)
1. Konformacijski osjetljivo protutijelo koje se može dobiti uz pomoć antigenskog konstrukta koji sadrži antigenski peptid koji ima aminokiselinski slijed β-amiloida, kojeg se bira iz skupine koju čine SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 i SEQ ID NO: 5, ili njegovog aktivnog fragmenta, gdje je antigenski peptid, ili njegov aktivni fragment, tako modificiran da ima kovalentno vezani polietilen-glikol, po jedan na svakom kraju, i rekonstituiran u liposomu, naznačen time što navedeno protutijelo ima specifičnost vezanja za antigenski peptid i
(a) pokazuje povećanu konformacijsku osjetljivost i afinitet za antigen u odnosu na protutijelo dobiveno uz pomoć palmitoiliranog antigenskog peptida; i
(b) potiče prijelaz β-nabranog lista u α-uzvojnicu amiloidnog peptida.1. A conformationally sensitive antibody that can be obtained with the help of an antigenic construct containing an antigenic peptide having the amino acid sequence of β-amyloid, selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, or its active fragment, where the antigenic peptide, or its active fragment, is modified to have covalently bound polyethylene glycol, one at each end, and reconstituted in a liposome, indicated by that said antibody has the specificity of binding to the antigenic peptide i
(a) shows increased conformational sensitivity and affinity for the antigen compared to an antibody raised with the aid of a palmitoylated antigenic peptide; and
(b) promotes the transition of the β-pleated sheet to the α-coil of the amyloid peptide.
2. Protutijelo u skladu s patentnim zahtjevom 1, naznačeno time što je navedeno protutijelo protutijelo izotipa IgG1.2. An antibody according to claim 1, characterized in that said antibody is an IgG1 isotype antibody.
3. Protutijelo u skladu s patentnim zahtjevom 1, naznačeno time što prilikom inkubiranja s amiloidnim vlaknima dovodi do vlakana veličine < 800 nm kod 40-60% od svih prisutnih vlakana.3. The antibody according to patent claim 1, characterized in that when incubated with amyloid fibers it leads to fibers < 800 nm in size in 40-60% of all fibers present.
4. Protutijelo u skladu s patentnim zahtjevom 1, dobiveno uz pomoć antigenskog konstrukta koji sadrži antigenski peptid koji ima aminokiselinski slijed SEQ ID NO: 1, tako modificiran da sadrži po jedan lizin kovalentno vezan na svakom kraju amiloidnog slijeda FRHDSGY, te polietilen-glikol (PEG) kovalentno vezan na lizin na jednom kraju i dioleil-fosfatidilkolin-etanolamin na drugom kraju molekule PEG, naznačeno time što navedeno protutijelo
(a) djelotvorno solubilizira vlakna Aβ1-40 i Aβ1-42; i
(b) štiti stanice PC12 in vitro od apoptoze i metaboličke inhibicije koju induciraju vlakna Aβ1-40 i Aβ1-42.4. Antibody according to patent claim 1, obtained with the help of an antigenic construct containing an antigenic peptide having the amino acid sequence SEQ ID NO: 1, so modified that it contains one lysine covalently bound at each end of the amyloid sequence FRHDSGY, and polyethylene glycol ( PEG) covalently bound to lysine at one end and diol-phosphatidylcholine-ethanolamine at the other end of the PEG molecule, indicated by the fact that the said antibody
(a) effectively solubilizes Aβ1-40 and Aβ1-42 fibers; and
(b) protects PC12 cells in vitro from apoptosis and metabolic inhibition induced by Aβ1-40 and Aβ1-42 fibrils.
5. Protutijelo u skladu s bilo kojim od prethodnih patentnih zahtjeva, naznačeno time što je navedeno protutijelo poliklonsko protutijelo ili monoklonsko protutijelo.5. An antibody according to any of the preceding patent claims, characterized in that said antibody is a polyclonal antibody or a monoclonal antibody.
6. Protutijelo u skladu s patentnim zahtjevom 4, naznačeno time što prilikom primjene na životinjskom ili ljudskom pacijentu dovodi do značajnih razina obnavljanja pamćenje i pobude znatiželje, bez izazivanja krvarenja u mozgu imuniziranog životinjskog ili ljudskog pacijenta.6. The antibody according to claim 4, characterized in that, when applied to an animal or human patient, it leads to significant levels of memory restoration and curiosity stimulation, without causing bleeding in the brain of the immunized animal or human patient.
7. Protutijelo u skladu s patentnim zahtjevom 1, naznačeno time što se veže na topljive oligomere Aβ1-40.7. The antibody according to claim 1, characterized in that it binds to soluble Aβ1-40 oligomers.
8. Upotreba epitopa kojeg se bira iz skupine koju čine SEQ ID NO: 2, SEQ ID NO: 3 i SEQ ID NO: 4, ili njegov aktivni fragment, uklopljen u antigenski konstrukt koji sadrži antigenski peptid, ili njegov aktivni fragment, tako modificiran da ima kovalentno vezani polietilen-glikol, po jedan na svakom kraju, i rekonstituiran u liposomu, naznačena time što je epitop namijenjen dobivanju protutijela u skladu s bilo kojim od patentnih zahtjeva 1-7.8. Use of an epitope selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, or its active fragment, incorporated into an antigenic construct containing an antigenic peptide, or its active fragment, thus modified to have covalently bound polyethylene-glycol, one at each end, and reconstituted in a liposome, characterized in that the epitope is intended for obtaining an antibody according to any of claims 1-7.
9. Protutijelo u skladu s bilo kojim od patentnih zahtjeva 1-7, naznačeno time što je namijenjeno pasivnom imuniziranju pojedinaca protiv neuroloških bolesti i poremećaja.9. An antibody according to any one of claims 1-7, characterized in that it is intended to passively immunize individuals against neurological diseases and disorders.
10. Protutijelo u skladu s patentnim zahtjevom 9, naznačeno time što je navedena bolest ili poremećaj Alzheimerova bolest.10. The antibody according to claim 9, characterized in that said disease or disorder is Alzheimer's disease.
11. Postupak proizvodnje konformacijski osjetljivog protutijela za pasivno imuniziranje pojedinaca protiv neuroloških bolesti i poremećaja, naznačen time što se navedeni postupak sastoji u rekonstituiranju antigenskog konstrukta u liposomima, gdje antigenski konstrukt sadrži peptid koji ima aminokiselinski slijed β-amiloida, ili njegovog aktivnog fragmenta, kojeg se bira iz skupine koju čine SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 i SEQ ID NO: 5, ili njegov aktivni fragment, gdje je antigenski peptid, ili njegov aktivni fragment, tako modificiran da ima kovalentno vezani polietilen-glikol, po jedan na svakom kraju, i potiče proizvodnju protutijela usmjerenog protiv antigenskog konstrukta.11. A process for the production of a conformationally sensitive antibody for passively immunizing individuals against neurological diseases and disorders, characterized in that the said process consists in the reconstitution of an antigenic construct in liposomes, where the antigenic construct contains a peptide that has the amino acid sequence of β-amyloid, or its active fragment, which is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, or its active fragment, where the antigenic peptide, or its active fragment , modified to have a covalently bound polyethylene glycol, one at each end, and induces the production of an antibody directed against the antigenic construct.
12. Postupak u skladu s patentnim zahtjevom 11, naznačen time što je neurološka bolest Alzheimerova bolest.12. The method according to claim 11, characterized in that the neurological disease is Alzheimer's disease.