CS247303B1 - Method of blood mixed immunoglobulin preparation overlay - Google Patents
Method of blood mixed immunoglobulin preparation overlay Download PDFInfo
- Publication number
- CS247303B1 CS247303B1 CS630184A CS630184A CS247303B1 CS 247303 B1 CS247303 B1 CS 247303B1 CS 630184 A CS630184 A CS 630184A CS 630184 A CS630184 A CS 630184A CS 247303 B1 CS247303 B1 CS 247303B1
- Authority
- CS
- Czechoslovakia
- Prior art keywords
- igm
- iga
- igg
- dialysis
- solution
- Prior art date
Links
- 108060003951 Immunoglobulin Proteins 0.000 title claims description 34
- 102000018358 immunoglobulin Human genes 0.000 title claims description 34
- 238000000034 method Methods 0.000 title claims description 15
- 239000008280 blood Substances 0.000 title claims description 3
- 210000004369 blood Anatomy 0.000 title claims description 3
- 238000002360 preparation method Methods 0.000 title description 20
- 238000000502 dialysis Methods 0.000 claims description 30
- 239000000243 solution Substances 0.000 claims description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 102000004169 proteins and genes Human genes 0.000 claims description 13
- 108090000623 proteins and genes Proteins 0.000 claims description 13
- 239000000126 substance Substances 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 7
- 230000001077 hypotensive effect Effects 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 7
- 239000002244 precipitate Substances 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 5
- 229940072221 immunoglobulins Drugs 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 4
- 208000001953 Hypotension Diseases 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 3
- 208000021822 hypotensive Diseases 0.000 claims description 3
- 238000000108 ultra-filtration Methods 0.000 claims description 3
- 229920000298 Cellophane Polymers 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 238000011146 sterile filtration Methods 0.000 claims description 2
- 238000005292 vacuum distillation Methods 0.000 claims description 2
- 229920002554 vinyl polymer Polymers 0.000 claims description 2
- 230000000521 hyperimmunizing effect Effects 0.000 claims 2
- 241000270322 Lepidosauria Species 0.000 claims 1
- 230000000844 anti-bacterial effect Effects 0.000 claims 1
- 230000001147 anti-toxic effect Effects 0.000 claims 1
- 230000000840 anti-viral effect Effects 0.000 claims 1
- 239000004922 lacquer Substances 0.000 claims 1
- 238000004062 sedimentation Methods 0.000 claims 1
- 210000002966 serum Anatomy 0.000 claims 1
- 230000003171 anti-complementary effect Effects 0.000 description 12
- 150000001875 compounds Chemical class 0.000 description 6
- 238000006722 reduction reaction Methods 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- 150000003384 small molecules Chemical class 0.000 description 6
- 108010074605 gamma-Globulins Proteins 0.000 description 5
- 230000009467 reduction Effects 0.000 description 5
- 239000013049 sediment Substances 0.000 description 5
- 102000004506 Blood Proteins Human genes 0.000 description 3
- 108010017384 Blood Proteins Proteins 0.000 description 3
- 230000036772 blood pressure Effects 0.000 description 3
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- NLHHRLWOUZZQLW-UHFFFAOYSA-N Acrylonitrile Chemical compound C=CC#N NLHHRLWOUZZQLW-UHFFFAOYSA-N 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- AYNSTGCNKVUQIL-UHFFFAOYSA-N C(CCCCCCCCCCC)C=1C=CC(=C(C=1)C1=NC(=CC(=C1)N(CCN(C)C)C)C1=C(C=CC(=C1)CCCCCCCCCCCC)OC)OC Chemical compound C(CCCCCCCCCCC)C=1C=CC(=C(C=1)C1=NC(=CC(=C1)N(CCN(C)C)C)C1=C(C=CC(=C1)CCCCCCCCCCCC)OC)OC AYNSTGCNKVUQIL-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000029936 alkylation Effects 0.000 description 2
- 238000005804 alkylation reaction Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000024203 complement activation Effects 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- VHJLVAABSRFDPM-ZXZARUISSA-N dithioerythritol Chemical compound SC[C@H](O)[C@H](O)CS VHJLVAABSRFDPM-ZXZARUISSA-N 0.000 description 2
- PGLTVOMIXTUURA-UHFFFAOYSA-N iodoacetamide Chemical compound NC(=O)CI PGLTVOMIXTUURA-UHFFFAOYSA-N 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 230000035488 systolic blood pressure Effects 0.000 description 2
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 230000009435 amidation Effects 0.000 description 1
- 238000007112 amidation reaction Methods 0.000 description 1
- 230000002391 anti-complement effect Effects 0.000 description 1
- 108010008730 anticomplement Proteins 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- AGEZXYOZHKGVCM-UHFFFAOYSA-N benzyl bromide Chemical compound BrCC1=CC=CC=C1 AGEZXYOZHKGVCM-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- -1 collodion Polymers 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 150000002484 inorganic compounds Chemical class 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000012792 lyophilization process Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 229940012957 plasmin Drugs 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000006337 proteolytic cleavage Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 229940078499 tricalcium phosphate Drugs 0.000 description 1
- 229910000391 tricalcium phosphate Inorganic materials 0.000 description 1
- 235000019731 tricalcium phosphate Nutrition 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Description
Nouvelle presse Medicale 5, 1193, 1976).Nouvelle Presse Medicale 5, 1193 (1976).
Súčasná technologická prax na jednej straně sice oddel'uje zmesný imunoglobulínový preparát na bázeOn the one hand, the current technological practice separates a mixed immunoglobulin preparation based on it
IgG 4- IgA 4- IgM od zmesi iných krvných bielkovín, ale na druhej straně zanáša do roztokuIgG 4-IgA 4-IgM from a mixture of other blood proteins, but on the other hand enters into solution
IgG 4- IgA 4- IgM nízkomolekulárné anorganické alebo organické zlúčeniny. Technologický postup može taktiež časť niektorých zlúčenín s biochemicko-fyziologickou aktivitou v roztoku zmesného imunoglobulínového preparátu koncentrovat, t. j. obohacovat vzhladom ku ich koncetrácii vo východzej surovině, z ktorej boli imunoglobulínyIgG 4-IgA 4-IgM low molecular weight inorganic or organic compounds. The technological process can also concentrate some of the compounds with biochemical-physiological activity in the solution of the mixed immunoglobulin preparation, i.e., enrich it due to their concentration in the starting material of which the immunoglobulins were
IgG 4- IgA + IgM izolované.IgG 4- IgA + IgM isolated.
Princip odstraňovania nízkomolekulárných látok s využitím dialýzy oproti destilovanej apyrogénnej vodě bod použitý u albuminu (Štěpánek I., Uhrík J.:_AO 208 866) ako aj u gama globulínu (Štěpánek I., AO číslo 244 306). Odstraňovanie nízkomolekulárnych látok s využitím dialýzy oproti 0,1% až 3,0 %-nérau vodnému roztoku chloridu sodného bolo použité u gama globulínu (Štěpánek I., Uhrík J.: AO 208 868) ako aj u zmesného imunoglobulínového preparátu na bázePrinciple of low molecular weight removal using dialysis against distilled pyrogen-free water the point used for albumin (Stepanek I., Uhrik J.:_AO 208 866) as well as gamma globulin (Stepanek I., AO No. 244 306). Removal of low molecular weight substances using dialysis versus 0.1% to 3.0% sodium chloride aqueous solution was used for gamma globulin (Stepanek I., Uhrik J .: AO 208 868) as well as a mixed immunoglobulin preparation based on
IgG + IgA + IgM (Štěpánek I., Uhrík ].: AO 208 867).IgG + IgA + IgM (Stepanek I., Uhrik] .: AO 208 867).
Zmesný imunoglobulínový preparát na bázeCompound based immunoglobulin preparation
IgG + IgA + IgM , připravený podl'a doposial' platných kritérií pre intramuskuiárné podanie má dokazatelný hypotenzívný a antikomplementárný účinok, čo je pre kvalitu intravenózného preparátu nepřípustné. Podanie preparátov týchto kvalit intravenózne može v dósledku náhlého poklesu krevného tlaku pacienta ohrozit a v dósledku antikomplementárnej aktivity oslabit jeho nešpecilickú obranu.IgG + IgA + IgM prepared according to the current criteria for intramuscular administration has a proven hypotensive and anticomplementary effect, which is unacceptable for the quality of the intravenous preparation. Administration of preparations of these qualities intravenously may compromise the patient's blood pressure as a result of a sudden drop in blood pressure and weaken his non-specific defense as a result of anticomplementary activity.
Na odstraňovanie antikomplementárnej aktivity gama globulínu, ktorý bol připravený zo zmesi plamzatických alebo sérových bielkovin, bola použitá úprava hodnoty pH na 4,0 pri 24 hodinovej inkubácii pri 37 CC (Barandun S., Kistler P., Jeunet F., Isliker H.: Vox Sang. 7, 157, 1962).To remove the anti-complementary activity of gamma globulin, which was prepared from a mixture of plasma or serum proteins, a pH adjustment of 4.0 was used for 24 hours incubation at 37 ° C (Barandun S., Kistler P., Jeunet F., Isliker H. Vox Sang., 7, 157 (1962).
Pokles antikomplementárnej aktivity gamaglobulínu bol zaznamenaný po přidaní aktívneho uhiia (Steinbuch M., Audran R., Amouch P., Blatrix C.: Vox Sang. 13, 103, 1967), po přidaní trikalciumfosfátu (Steinbuch M., Audran R., Amouch P., Blatrix C.: Rev. med. Tours, suppl. No 3, 79, 1968) a po přidaní albuminu (Auerswald W., Kiese wetter E.: Wien. med. Wschr. 115, 690,1965).A decrease in the anticomplementary activity of gamaglobulin was observed after the addition of activated carbon (Steinbuch, M., Audran, R., Amouch, P., Blatrix, C .: Vox Sang. 13, 103, 1967), after the addition of tricalcium phosphate (Steinbuch, M., Audran, R., Amouch. P., Blatrix C .: Rev. Med Tours, Suppl. No 3, 79, 1968) and after the addition of albumin (Auerswald W., Kiese Wetter E .: Wien. Med. Wschr. 115, 690, 1965).
Znižovanie antikomplementárnej aktivity gama globulínového roztoku pomocou proteolytického štiepenia bolo realizované pepsínom (Schultze Η. E., Schwick G.: Dtsch. med. Wschr. 87, 1643, 1962) a plazmínom (Sgouris J. T.: Vox Sang. 13, 71, 1967).Reduction of the anti-complementary activity of the gamma globulin solution by proteolytic cleavage was realized by pepsin (Schultze, E., Schwick, G .: Dtsch. Med. Wschr. 87, 1643, 1962) and plasmin (Sgouris JT: Vox Sang. 13, 71, 1967). .
Antikomplementárnu aktivitu gama globulínovej molekuly je možné znížiť jej chemickou redukciou beta merkaptoetanolom (Wiederman G., Miescher P. A., Frankiin E. C.: Proč. Soc. exp. Biol. Med. 113, 609, 1963), modifikáciou betapropiolaktonoin (Stephan W.: Z. kliň. Chem. 7, 282, 1969; Stephan W.: Vox Sang. 28, 422, 1975), redukciou s dithioerythritolom (Isenman D. E. Dorrington K. )., Painter R. H.: J. Immun. 114, 1726, 1975), sulfonáciou (Masuho Y., Tomibe K., Matsuzawa K., Ohtsu A.: Vox Sang. 32, 175, 1977), amidáciou (Schmidtberger R.: US Patent No 4 118 379 [1978)), redukciou s dithioethreitolom a alkyláciou s jodacetamidom (Fernandes Ρ. M., Lundhlad J. I,.: Vox Sang. 39, 101, 1980) a redukciou s dithiothreitolom v kombinácii s dithioerythritolom s následnou alkyláciou pomocou jodacetamidu alebo metyljodidu, akrylonitrilu, benzylbromidu, připadne etylénoxidu (Schroeder D. D., Tankersiey D. L., Lundblad J. L.: Vox Sang. 40, 373, 1981).The anti-complementary activity of the gamma globulin molecule can be reduced by its chemical reduction by beta mercaptoethanol (Wiederman G., Miescher PA, Frankiin EC: Proc. Soc. Exp. Biol. Med. 113, 609, 1963), by modification of betapropiolactonoin (Stephan W .: Z. Chem., 7, 282 (1969), Stephan W .: Vox Sang, 28, 422, 1975), reduction with dithioerythritol (Isenman DE Dorrington, K.), Painter RH: J. Immun. 114, 1726, 1975), by sulfonation (Masuho Y., Tomibe K., Matsuzawa K., Ohtsu A .: Vox Sang. 32, 175, 1977), by amidation (Schmidtberger R .: US Patent No 4,118,379 [1978] ), reduction with dithioethreitol and alkylation with iodoacetamide (Fernandes, M., Lundhlad J.I., Vox Sang. 39, 101, 1980) and reduction with dithiothreitol in combination with dithioerythritol followed by alkylation with iodoacetamide, methyl iodide, acrylonitrile, methyl iodide, acrylonitrile, methyl iodide, benzyl bromide and ethylene oxide, respectively (Schroeder DD, Tankersiey DL, Lundblad JL: Vox Sang. 40, 373, 1981).
Antikomplementárná aktivita zmesného imunoglobulínového preparátu na bázeAnti-complementary activity of a mixed immunoglobulin preparation based on
IgG + IgA + IgM súvisí pravděpodobně so vznikom polymérov IgG + IgA + IgM , ktoré můžu vznikat v procese technologickej izolácie a jednak v procese lyofilizácie ako dosledok interakcie nízkomolekulárných látok s molekulami IgG, IgA a IgM.IgG + IgA + IgM is likely to be associated with the formation of IgG + IgA + IgM polymers, which may arise in the process of technological isolation and in the lyophilization process as a result of the interaction of low molecular weight substances with IgG, IgA and IgM molecules.
Použitie destilovanej apyrogénnej vody pri separácii nizkomolekulárnych zlúčenín zo zmesného imunoglobulínového preparátu na bázeUse of distilled pyrogen-free water in the separation of low molecular weight compounds from a mixed immunoglobulin preparation based on
IgG -i- IgA + IgM sposobuje precipitáciu pravděpodobně polymérov IgG, IgA a IgM a jednak odstránenie nizkomolekulárnych látok, do ktorých můžeme zahrnúť aj zlúčeniny s hypotenzívnou aktivitou, v důsledku čoho pri lyofilizácii nemůže dojsť ku ich interakcii s molekulami IgG, IgA a IgM, čiže nedochádza ani ku polymerizácii, to znamená, že ani ku vzniku antikomplementárnej aktivity.IgG-IgA + IgM causes the precipitation of IgG, IgA and IgM polymers and the elimination of low molecular weight substances, which may include compounds with hypotensive activity, which, as a result of lyophilization, cannot interact with IgG, IgA and IgM molecules, thus, there is no polymerization, i.e. no anti-complementary activity.
Antikomplementárná aktivita bcla testovaná na základe úbytku aktivity komplementu. Úbytok známej hodnoty aktivity komplementu po přidaní známého množstva roztoku lyofilizovaného zmesného imunoglobulínového preparátu na bázeAnti-complement activity was tested for complement activity loss. Loss of known value of complement activity after addition of known amount of lyophilized mixed immunoglobulin preparation based on solution
IgG + IgA + IgM bol stanovený metodou podlá Kabáta a Mayera (Kabát E. A., Mayer Μ. M.: Experimentální Imunochemie Praha ČSAV 1965 str. 180).IgG + IgA + IgM was determined by the method of Kabat and Mayer (Kabat E.A., Mayer, M.: Experimental Immunochemistry Prague, Czechoslovak Academy of Sciences 1965, p. 180).
Efekt destilovanej apyrogénnej vody pri separácii antikomplementárnej aktivity je možné dokumentovat u 10 experimentálných šarží zmesného imunoglobulínového preparátu na bázeThe effect of distilled pyrogen-free water in the separation of anticomplementary activity can be documented in 10 experimental batches of a mixed immunoglobulin preparation based on
IgG + IgA -i- IgM , kde bolo dosiahnuté zníženie antikomplementárnej aktivity u lyofilizovaného zmesného imunoglobulínového preparátu na báze IgG + IgA + IgM o 90 až 98 % v porovnaní s tým istým materiálom, ktorý bol lyofilizovaný bez predchádzajúcej separácie nízkomolekulárných zlúčenín.IgG + IgA-IgM, where a reduction in anticomplementary activity of the lyophilized mixed IgG + IgA + IgM-based immunoglobulin preparation was achieved by 90-98% compared to the same material that was lyophilized without prior separation of the low molecular weight compounds.
Hypotenzívny účinok bol testovaný na základe poklesu krvného tlaku králika priamou krvnou metodou podlá ČSL 3 (ČSL 3, svazok I, vydanie III, Avicenum Praha str. 149) s tým, že králíkovi o hmotnosti 2 až 3 kg sa aplikujú 2 ml zmesného imunoglobulínového preparátu na bázeThe hypotensive effect was tested by a decrease in rabbit blood pressure by a direct blood method according to CSL 3 (CSL 3, Volume I, Edition III, Avicenum Praha p. 149) with 2 ml of a mixed immunoglobulin preparation being administered to a rabbit weighing 2-3 kg. based
IgG + IgA + IgM na kg hmotnosti. Obsah bielkovin v aplikovanom zmesnom imunoglobulínovom preparáte na bázeIgG + IgA + IgM per kg of weight. Protein content of the applied mixed immunoglobulin preparation
IgG + IgA t IgM je 15 g na 1 090 ml roztoku. U preparátov připravených podlá navrhovaného postupu sa hypotenzívná aktivita nevyskytovala v tak výraznej intenzitě, čo je možné dokumentovat údajom, že z 10 připravených experimentálnych šarží podlá předloženého vynálezu bol stanovený pokles systolického tlaku maximálně o 4 až 8 % v porovnaní s tým istým materiálom v tej istej dávke bez separácie nízkomolekulárných zlúčenín, kde bol stanovený pokles systolického tlaku od 20 do 30 %.IgG + IgA t IgM is 15 g per 1090 ml solution. In the formulations prepared according to the proposed procedure, the hypotensive activity did not occur to such an appreciable intensity, as evidenced by the data that from a maximum of 10 experimental batches of the present invention a systolic pressure drop of at most 4 to 8% was determined compared to the same material. dose without separation of low molecular weight compounds, where the systolic pressure drop was determined to be from 20 to 30%.
Podstatou vynálezu je odstránenie nízkomolekulárných zlúčenín zo zmesného roztokuIt is an object of the present invention to remove low molecular weight compounds from the mixed solution
IgG + IgA + IgM prietokovou dialýzou oproti destilovanej apyrogénnej vodě, pričom pri použití cca 300 násobného objemu vody ako je objem dialyzovaného roztoku dojde ku precipitácii časti imunoglobulínov, pravděpodobně po^mérných foriem IgG, IgA a IgM, ktoré vyprecipitujú v ipodobe sedimentu a predstavujú zdroj IgM. Roztok po -oddělení sedimentu představuje zmesný imunoglobulínový preparát na bázeIgG + IgA + IgM by flow dialysis versus distilled pyrogen-free water, using about 300 times the volume of water such as the volume of the dialyzed solution precipitates a portion of the immunoglobulins, probably the proportional forms of IgG, IgA and IgM, which precipitate similar to sediment IgM. The sediment separation solution is a mixed immunoglobulin preparation based on
IgG + IgA + IgM so zníženým hypotenzívnym a antikomplementárnym účinkom.IgG + IgA + IgM with reduced hypotensive and anti-complementary effect.
Bielkovinný materiál obsahujúci purifikovanýProtein material containing purified
IgG + IgA + IgM sa suspenduje za aseptických podmienok v takom množstve apyrogénnej destilovanej vody o teplote 0 °C až +6 °C, aby hodnota koncentrácie bielkovin sa nachádzala v rozmedzí 1 θ/o až 15 %, s optimom 8 + 2 %, roztok imunoglobulínov sa vyčíri, například filtráciou a separácia nízkomolekulárných zlúčenín sa prevádza z roztoku imunoglobulínov prietokovou dialýzou oproti destilovanej apyrogénnej vodě s hodnotou pH v rozmedzí 4,0 až 8,0 pri prietoku 300 ml až 2 000 ml za minútu po dobu 24 až 72 hodin.IgG + IgA + IgM is suspended under aseptic conditions in an amount of pyrogen-free distilled water at a temperature of 0 ° C to + 6 ° C such that the protein concentration is in the range of 1 θ / o to 15%, with an optimum of 8 + 2%. the immunoglobulin solution becomes clear, for example by filtration, and the separation of the low molecular weight compounds is carried out from the immunoglobulin solution by flow dialysis versus distilled pyrogen-free water at a pH ranging from 4.0 to 8.0 at a flow rate of 300 ml to 2000 ml per minute for 24 to 72 hours .
Po ukončení dialýzy sa z dialyzačného potrubia oddialyzovaný roztok imunoglobulínov vyleje a vzniklý sediment sa odstráni například Siltráciou alebo centrifugáciou. Roztok imunoglobulínov sa připadne zakoncentrováva ultrafiltráciou, podrobí sa sterilizácii například filtráciou alebo radiačnou sterfizáciou,. rozplňuje sa, lyofilizuje a v případe potřeby sa podrobí ďalšiemu spracovaniu.After dialysis, the dialyzed immunoglobulin solution is poured from the dialysis line and the resulting sediment is removed, for example, by centrifugation or centrifugation. The immunoglobulin solution is optionally concentrated by ultrafiltration, subjected to sterilization, for example, by filtration or radiation sterilization. it is filled, lyophilized and, if necessary, subjected to further processing.
Vynález je ďalej objasněný na príkladoch prevedenia, ktorými jeho rozsah nie je ani obmedzený ani vyčerpaný.The invention is further elucidated by means of exemplary embodiments in which its scope is neither limited nor exhausted.
Příklady prevedeniaExamples of design
Příklad 1Example 1
Východzou surovinou može byť bielkovinná pasta zmesného imunoglobulínového preparátu na bázeThe starting material may be a protein paste of a mixed immunoglobulin preparation based on it
IgG + IgA 4- IgM získaná například sposobom podlá Pejaudiera a spol. [Pejaudier L., Audran R., Steinbuch M.: Vox Sang. 23, 165, 1972], Bielkovinný materiál vyprecipitovaných imunoglobulínov sa rozpúšfa v takom množstve destilovanej apyrogénnej vody o teplote 0 °C až +6 °C, aby hodnota koncentrácie bielkovín sa nachádzala v rozmedzí 1 až 15 %, s optimom 8 + 2 %. Rozpúštanie sa urýchluje miešaním. Po rozpuštění sa roztok imunoglobulínov na bázeIgG + IgA 4- IgM obtained, for example, by the method of Pejaudier et al. [Pejaudier L., Audran R., Steinbuch M .: Vox Sang. 23, 165, 1972], The protein material of the precipitated immunoglobulins is dissolved in an amount of distilled pyrogen-free water at a temperature of 0 ° C to + 6 ° C such that the protein concentration is in the range of 1 to 15%, with an optimum of 8 + 2%. Dissolution is accelerated by stirring. After dissolution, the solution of immunoglobulins is based on
IgG + IgA + IgM vyčíri filtráciou.IgG + IgA + IgM is clarified by filtration.
V technologických pomeroch sú vyššie uvedené podmienky spravidla zachované vtedy, keď rozpúšťame 1 kg bielkovinnej pasty imunoglybulínov skupinyIn technological conditions, the above conditions are generally maintained when dissolving 1 kg of immunoglybulin group protein paste
IgG + IgA + IgM v '2 000 ml apyrogénnej destilovanej vody o teplote 0°C až +6°C. Hodnota pH takto vzniknutého roztoku imunoglobulínov bývá spravidla medzi 4,0 až 8,0 a koncentrácia bielkovin v rozmedzí 8 + 2 °/o. Takto starostlivo připravený roztok sa vyčíri například filtráciou a nízkomolekulárné zlúčeniny sa odstránia z roztoku prietokovou dialýzou cez dialyzačné potrubie. Dialyzačné potrubie može být vyrobené z celofánových, kolódiových, vinylpolymérnych membrán, připadne z iných materiálov.IgG + IgA + IgM in 2000 ml of pyrogen-free distilled water at 0 ° C to + 6 ° C. The pH of the resulting immunoglobulin solution is generally between 4.0 and 8.0 and the protein concentration is in the range of 8 + 2%. The carefully prepared solution is clarified, for example, by filtration, and the low molecular weight compounds are removed from the solution by flow dialysis through a dialysis line. The dialysis tubing may be made of cellophane, collodion, vinyl polymer membranes, or other materials.
Namočené dialyzačné potrubie v apyrogénnej destilovanej vodě sa nechá nabobtnať a potom sa naplní destilovanou apyrogénnou vodou za účelom preverenia celistvosti. Priemer dialyzačného potrubia by sa mal pohybovat od 2 cm do 15 cm s optimom· 5 cm. Sterilizácla dialyzačného potrubia sa prevádza v roztoku destilovanej apyrogénnej vody, například autoklavovaním pri 122 °C po dobu 40 minút, alebo chemickou cestou, například etylénoxidom. Spodná časť dialyzačného potrubia sa uzavrie uzlom. Roztok zmesného imunoglobulínového preparátu na bázeThe soaked dialysis tubing in pyrogen-free distilled water is swollen and then filled with distilled pyrogen-free water to verify integrity. The diameter of the dialysis pipe should be between 2 cm and 15 cm with an optimum of 5 cm. The dialysis line sterilization is carried out in a solution of distilled pyrogen-free water, for example by autoclaving at 122 ° C for 40 minutes, or by chemical means, for example ethylene oxide. The lower part of the dialysis line is closed with a knot. Mixed immunoglobulin based solution
IgG + IgA + IgM , sterilizovaný například filtráciou, sa naleje do vysterilizovaného dialyzačného potrubia, uzatvoreného v spodnej časti uzl-om a uzatvori sa vo vrchnej časti uzlom so slučkou, za ktorú sa zavěsí na vrchné víko dialyzačnej prietokovej kolóny.IgG + IgA + IgM, for example, sterilized by filtration, is poured into a sterilized dialysis tubing, closed at the bottom with a node, and closed at the top with a loop knot, after which it is suspended on the top of the dialysis flow column.
Za optimálně rozměry technologických dialyzačné prietokových kolon sa považujú také, kde priemer kruhovej alebo uhlopriečka štvorcovej, připadne obdížnikovej základnej plochy ku výške kolóny sa pohybuje v pomere 1 : 5 až 1 : 10. Množstvo roztoku zmesného imunoglobulínového preparátu vloženého do dialýzy nemá překročit 25 až 30 % objemu prázdnej kolóny.The optimum dimensions of the technological dialysis flow columns are those where the diameter of the circular or diagonal of the square or rectangular base to the height of the column is in the ratio of 1: 5 to 1: 10. The amount of mixed immunoglobulin preparation inserted into dialysis should not exceed 25 to 30 % of the volume of the empty column.
Po nanesení zvoleného objemu roztoku zmesného imunoglobulínového preparátu na bázeAfter application of the selected volume of solution of the mixed immunoglobulin preparation on a base
IgG + IgA + IgM sa zahájí nútený prietok apyrogénnej destilovanej vody o teplote 0 CC až +6 °C o pH 4,0 až 8,0 pri prietoku 300 ml až 2 000 ml za minutu, s výhodou pri prietoku 1 000 ml za minútu po dobu 72 hodin. Množstvo celkové pretečenej destilovanej apyrogénnej vody má byť minimálně 300 násobkom objemu zmesného imunoglobulínového roztoku, ktorý bol umiestnený do vnútra dialyzačného potrubia.IgG + IgA + IgM is initiated with a forced flow of pyrogen-free distilled water at a temperature of 0 ° C to + 6 ° C at a pH of 4.0 to 8.0 at a flow rate of 300 ml to 2000 ml per minute, preferably at a flow rate of 1000 ml. minute for 72 hours. The amount of total flowed distilled pyrogen-free water should be at least 300 times the volume of the mixed immunoglobulin solution that was placed inside the dialysis line.
Pri penetrácii nízkomolekulárných zlúčenín z roztoku zmesného imunoglobullnového preparátu cez dialyzačnú membránu odchádzajú aj nízkomolekulárné zlúčeniny sposobujúce hypotenzívny účinok v dósledku odchodu nízkomolekulárných zlúčenín cez stenu dialyzačnej membrány klesá stabilita a tým aj rozpustnost, pravděpodobně časti polymcrov IgG, IgA a IgM, ktoré začnú vypadávat v podobě suspenzie až seílimentačného precipitátu a z roztoku spravidla sa usadila v dolnej časti potrubia. Se diment precipitátu sa spravidla objaví cca po 40 hodině priebehu dialýzy. Nachádzajú sa v ňom bielkoviny, ktoré sú zodpovědné za prevažnú časť antikomplementárnej aktivity.When penetrating the low-molecular compounds from the mixed immunoglobulin preparation solution through the dialysis membrane, also the low-molecular compounds exerting a hypotensive effect leave the dialysis membrane as a result of leaving the low-molecular compounds through the dialysis membrane wall. As a rule, it settled in the lower part of the pipe. The precipitate precipitate usually appears after about 40 hours of dialysis. It contains proteins that are responsible for most of the anticomplementary activity.
Po skončení dialýzy sa dialyzačné potrubie zavěsí so závěsného háčika na vrchnnm veku dialyzačnéj kolony, opatrné sa vyberie z kolony a po rozstrihnutí sa roztok oddělí od sedimentu a podrobí sa sterilnej filtrácii, pričom dojde ku definitívnemu oddeleniu zbytkov sedimentu z roztoku, roztok sa podrobí zakoncentrovaniu pomocou ultrafiltrácie, vymrazovaním, alebo vákuovej destilácii, rozplňuje sa a lyofilizuje a v případe potřeby sa podrobí ďalšiemu spracovaniu.When dialysis is complete, the dialysis tubing is suspended from the upper hook of the dialysis column, carefully removed from the column and, after shearing, the solution is separated from the sediment and subjected to sterile filtration, finally settling the sediment residue from the solution, concentrating the solution with by ultrafiltration, freeze-drying or vacuum distillation, is filled and lyophilized and, if necessary, subjected to further processing.
Příklad 2Example 2
Východziou surovinou može byt bielkovinná pasta zmesného imunoglobulínového preparátuThe starting material may be a protein paste of a mixed immunoglobulin preparation
IgG 4- IgA 4- IgM alebo ich roztok izolovaný aj pomocou iných metod. Bielkovinná pasta alebo roztok imunoglobulínov na bázeIgG 4-IgA 4-IgM or a solution thereof also isolated by other methods. Protein paste or immunoglobulin solution based on
IgG + IgA 4- IgM sa spracováva v dalšom postupe ako v příklade č. 1.IgG + IgA 4-IgM is processed in a further procedure as in Example 1.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS630184A CS247303B1 (en) | 1984-08-21 | 1984-08-21 | Method of blood mixed immunoglobulin preparation overlay |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS630184A CS247303B1 (en) | 1984-08-21 | 1984-08-21 | Method of blood mixed immunoglobulin preparation overlay |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CS247303B1 true CS247303B1 (en) | 1986-12-18 |
Family
ID=5409735
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS630184A CS247303B1 (en) | 1984-08-21 | 1984-08-21 | Method of blood mixed immunoglobulin preparation overlay |
Country Status (1)
| Country | Link |
|---|---|
| CS (1) | CS247303B1 (en) |
-
1984
- 1984-08-21 CS CS630184A patent/CS247303B1/en unknown
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4216205A (en) | Process of preparing a serum protein composition for intravenous application | |
| EP0073371B1 (en) | Intravenously injectable immune serum globulin and method of preparing same | |
| US4499073A (en) | Intravenously injectable immune serum globulin | |
| US5164487A (en) | Manufacturing intravenous tolerable immunoglobulin-g preparation | |
| RU2612899C2 (en) | Method for preparing composition of immunoglobulins | |
| RU2008916C1 (en) | Method for aqueous solution of immunoglobulin pasteurization | |
| SK18662000A3 (en) | Process for producing immunoglobulins for intravenous administration and other immunoglobulin products | |
| CN106459140B (en) | Method for purifying immunoglobulins | |
| SK287633B6 (en) | Method for the production of human gammaglobulin G and virus-inactivated human gammaglobulin G | |
| US4017360A (en) | Method for purifying hepatitis B antigen | |
| US5041537A (en) | Method of preparing a high-purity, virus safe, biologically active transferrin preparation | |
| EP0764447B1 (en) | Preparation of virally inactivated intravenously injectable immune serum globulin | |
| JPH0365327B2 (en) | ||
| Fernandes et al. | Preparation of a stable intravenous gamma‐globulin: process design and scale‐up | |
| FI61191C (en) | FOERFARANDE FOER UTVINNING AV IMMUNOGLOBULIN FOER INTRAVENOES ADMINISTRATION | |
| CA2253300A1 (en) | Biological material free of viral and molecular pathogens and a process for the production thereof | |
| US10358462B2 (en) | Method for the preparation of immunoglobulins | |
| CS247303B1 (en) | Method of blood mixed immunoglobulin preparation overlay | |
| CA1168152A (en) | Process for preparing human plasma fractions containing immune globulin (igg) | |
| AU2016231646B2 (en) | Method for the preparation of immunoglobulins | |
| CA2943328C (en) | Method for the preparation of immunoglobulins | |
| CS266252B1 (en) | Process for preparing blood mixed preparate on base igg +iga+igm | |
| CS244306B1 (en) | Blood gamma globulin preparation method with reduced hypotensive and anti complementary effect | |
| Ng et al. | Process‐Scale Purification of Immunoglobulin M Concentrate | |
| JPS5840930B2 (en) | Manufacturing method for hepatitis B vaccine |