CN1454901A - Gynaecological anti-infective specificity IgY and its combined preparation - Google Patents
Gynaecological anti-infective specificity IgY and its combined preparation Download PDFInfo
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- CN1454901A CN1454901A CN 03136958 CN03136958A CN1454901A CN 1454901 A CN1454901 A CN 1454901A CN 03136958 CN03136958 CN 03136958 CN 03136958 A CN03136958 A CN 03136958A CN 1454901 A CN1454901 A CN 1454901A
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Abstract
The present invention discloses a specific IgY for resisting gynecological infection, its preparation method and preparation. Said preparation method includes screening out the pathogenic bacteria resulted in gynecological infection diseases, on the basis of screening and sort-merging various pathogenic bacteria respectively preparing single or composite pathogenic bacteria antigen, preparing immune egg, preparing specific composite IgY crude product for resisting gynecological infection pathogenic bacteria and purifying said crude product. Said several preparations can be used for curing woman bacterial, mycotic, trichomonal and senile vaginitis or their mixed infection, and can obtain reliable therapeutic effect.
Description
Technical field
The present invention relates to a kind of anti-gynecological infection specific IgY and preparation method thereof, particularly relate to and be used to prevent and treat various gynecological infectious diseases specificity composite IgYs and preparation method thereof and combination preparation
Background technology
Current domestic and international treatment gynaecology trichomonal vaginitis, bacillary (non-specific) vaginitis, colpitis mycotica and their infectious diseases such as polyinfection are mainly used control infection such as chemostefilant, antibiotic and herbal medicine, diminish inflammation.Though these methods have certain effect, also there are problems, the lavation of life-time service chemostefilant, hip bath or insert cause the epidermis mucosa injury easily, as irritated, red and swollen, dry and cracked, in addition hemorrhage.Then cause bacterial drug resistance easily with antibiotic therapy, the life-time service broad-spectrum antibiotics also causes superinfection easily.Medical matters circle cause fungal infection with antibacterium medicine treatment vaginitis (non-specific) in a large number, with antifungals such as clotrimazole treatment colpitis mycotica, cause infectation of bacteria, and this class example can be found everywhere.Though adopt Chinese herbal and crude drugs preparations that certain effect is arranged, effect is also not obvious.
The more important thing is in applied chemistry medicine, the above-mentioned gynecological infection inflammatory process of antibiotic therapy, can destroy the local microecological balance of vaginal environment, not only killed harmful bacterium, also killed normal vaginal microbial flora, as probioticss such as lactobacillus, coryneform bacteria, faecalis, non-Streptococcus hemolyticuss.Lactobacillus makes local weak acid environment and the low pH value of forming of vagina, thereby hypertrophy and the adhesion of other microorganisms have been suppressed, other bacillus breeding has promoted useful anerobe growth again, thereby has kept vagina normal microflora balance from the pros and cons, and is very useful to increasing vagina immunizing power.Use pharmaceutical chemicals and antibiotic therapy gynecological inflammation, then will directly kill probioticss such as lactobacillus, destroy vagina normal microflora balance, the barrier of the original natural defence disease of forfeiture vagina increases the weight of the state of an illness on the contrary.Therefore at present both at home and abroad to this multiple common disease of gynecological infection, still there are not ideal, medicine green, that have no side effect.
Anti-gynecological infectious diseases specificity composite yolk immune antibody (Immunoglobulin of yolk IgY) belongs to the IgG immunoglobulin like protein, it is multiple pathogenic agent with gynecological infectious diseases, immunity breeding hen, make it to produce a large amount of corresponding immunoglobulin (Ig)s, through the extraction separation purifying, and make specific composite yolk immune antibody.These compound antibodies have specificity in conjunction with restraining effect to causing the pathogenic micro-organism of multiple gynecology colpitis, and (tens of ng/ml) just can play useful effect under the extremely low situation of concentration.Its feature is only the harmful bacterium of vagina to be played killing action, and to the symbiosis probiotics, and is then like water off a duck's back, and this and pharmaceutical chemicals and antibiotic are distinct.With this compound antibody (IgY) treatment, can protect the vaginal environment microecological balance, play and not only cure the disease but also function of health care.This treats the gynecological infectious diseases aspect at home and abroad, is an initiative precedent.
Technology contents
The purpose of this invention is to provide the specificity composite IgY of the multiple pathogenic agent of the common gynecological infectious diseases of a kind of direct inhibition and the preparation method of preparation thereof; The present invention also aims to provide a kind of by can directly pressing down the multiple colpitic different preparations of control that natural, safety that the specific IgY that kills the common vaginitis pathogenic agent of gynaecology makes have specific action again.
The objective of the invention is to be achieved through the following technical solutions:
(1) screening causes the germ of gynecological infectious diseases: vagina Gardner bacillus, Candida albicans, bacillus ducreyi, Trichomonas vaginalis etc." antigenic cross property " according to these representational pathogenic bacterium carries out sort merge.
(2) on the basis of screening and the various gynecological infection pathogenic bacterium of sort merge, prepare single respectively or compound gynecological infection pathogenic bacterium antigen:
According to cultural characters and the optimal culture conditions of various gynecological infection pathogenic bacterium, select the suitable culture base respectively, carry out the strain of bacterium worm and breed in a large number, collect culture, purification, bacterial strain counting, dilution packing;
With various single or blended gynecological infection pathogenic bacterium adding freund adjuvants, put into the high-speed homogenization machine again, the high speed pulverization homogenize forms milk-like slurry, makes single or compound gynecological infection pathogenic bacterium antigen.
(3) prepare immune egg
Selection has (SPF) healthy bird inlay that the healthy no-special pathogen that produces facing out of high immunne response ability or laying eggs carries, every at chest muscle or subcutaneously inject prepared single or compound gynecological infection pathogenic bacterium antigen respectively, the interval is 7 days after injection for the first time, again for the second time with same dosage, method injection, injected the 3rd time with same dose in 7 days at interval again after the injection for the second time, the injection back was chosen since the 20th day and is got high immune egg for the first time, and corresponding different antigen, the immune egg of being searched is carried out coded markings.
(4) preparation of anti-gynecological infection pathogenic bacterium specificity composite IgY crude product
With synantigen not respectively the hen of immunity produce the egg balanced mix, with 0.5% benzalkonium bromide solution or 0.1%KMnO4 solution or other similar thimerosals immune egg is soaked sterilization in 15-30 minute, the sterile distilled water flushing, dry, place beating machiae that eggshell is smashed, stir except that egg white stays 4-8 times of distilled water diluting of yolk adding with the yolk sieving, regulate 4-6 ℃ of following standing over night of PH to 5.5-6.0 with 1mol/lNaOH liquid or 1mol/l HCL liquid, diluent 1000-3000r/min high speed centrifugation 20 minutes, get supernatant liquor and hold back ultrafiltration and concentration 10-20 doubly with 170,000 molecular weight to 20, ten thousand molecular weight, concentrated solution is done promptly to get with the spray of cryospray desiccating method and is resisted gynecological infection specificity composite IgY crude product.
(5) the pure product preparation of anti-gynecological infection pathogenic bacterium specificity composite IgY
Adopt the disclosed patented technology (number of patent application 00101270.3 of our company, patent publication No. 1307061) crosses ion exchange column and gel exchange column respectively the anti-gynecological infection composite IgY crude product of producing by above technology is carried out purifying, promptly get pure composite IgY;
Use SDS-PAGE cataphoretic determination method, the anti-gynecological infection pathogenic bacterium specific IgY crude product of producing by above technology is detected, recording wherein, composite IgY content is more than 50%.
(6) adopt euzymelinked immunosorbent assay (ELISA) (ELISE) record the composite IgY biological activity tire reach 1: 256~512.
This anti-gynecological infection pathogenic bacterium specific IgY of 0.02-30% is equipped with the auxiliary material of 99.98-70%, can be made into various clinical acceptable forms, as washing lotion, sprays, effervescent (effervescent tablet, effervesce capsule), gelifying agent, ointment, suppository etc., be used to prevent and treat common gynecological infectious diseases.
The present invention directly imposes on vulva, vagina, the most concentrated position of uterine cervix pathogenic agent with above formulation, directly kill, effective, instant effect, solved the routine administration mode and made medicine enter the various bad drawback that is produced after the human circulation, also overcome conventional medicine and used vaginal mucosa drying, redness, allergy or the bleeding that the back is caused.The present invention is a specific specificity, fully natural green biotechnological formulation; vagina symbiosis probiotics there is good provide protection; can not resemble fully and pathogenic bacterium and probiotics be killed together administration of antibiotics or the chemical disinfection medicine, destroy the vaginal environment microecological balance, or cause resistance.Reached and not only cured the disease but also the dual function of prevention and health care.
Experimental example 1: vagina Gardner bacillus 10-20 hundred million, Trichomonas vaginalis purifying antigen protein 10 0ug are made 0.5ml bacterium liquid, add the abundant mixing of 0.5ml freund adjuvant, make complex antigen, use this antigen immune hen again, prepare anti-vagina Gardner bacillus, Trichomonas vaginalis specificity composite IgY.Then, serve as to detect antigen with vagina Gardner bacillus, Trichomonas vaginalis, vagina Candida albicans, bacillus ducreyi, detect prepared anti-gynecological infection specificity composite IgY antibodies with " ELISA method " (enzyme linked immunosorbent assay) and tire.Result such as following table
????IgY | Detection antigen | Antibodies is tired |
Anti-vagina Gardner bacillus, Trichomonas vaginalis specificity composite IgY | Vagina Gardner bacillus | ????1∶512 |
Trichomonas vaginalis | ????1∶512 | |
The vagina Candida albicans | ????1∶256 | |
Bacillus ducreyi | ????1∶256 |
Find out from above detected result, adopt method of the present invention to produce pathogenic bacterium antigen, adopt reinforced immunological method immunity bird inlay of the present invention again, prepared anti-vagina Gardner bacillus, Trichomonas vaginalis specificity composite IgY have the ideal broad spectrum, not only vagina Gardner bacillus, the Trichomonas vaginalis antigen to direct immunization has very strong restraining effect, the antibodies of being surveyed tire higher (1: 512); Other two kinds of gynecological infection pathogenic bacterium that do not have direct immunization are had certain restraining effect too, and detected antibodies is tired and is still belonged to ideal.
Experimental example 2: vagina Candida albicans 10-20 hundred million and bacillus ducreyi 10-20 hundred million are made 0.5ml bacterium liquid, add the abundant mixing of 0.5ml freund adjuvant, make complex antigen, use this antigen immune hen again, prepare anti-vagina Candida albicans, bacillus ducreyi specificity composite IgY.Then, serve as to detect antigen with vagina Gardner bacillus, Trichomonas vaginalis, vagina Candida albicans, bacillus ducreyi, detect prepared anti-gynecological infection specificity composite IgY antibodies with " ELISA method " (enzyme linked immunosorbent assay) and tire.Result such as following table
????IgY | Detection antigen | Antibodies is tired |
Anti-vagina Candida albicans, bacillus ducreyi specificity composite IgY | Vagina Gardner bacillus | ????1∶256 |
Trichomonas vaginalis | ????1∶256 | |
The vagina Candida albicans | ????1∶512 | |
Bacillus ducreyi | ????1∶512 |
Find out from above detected result, adopt method of the present invention to produce pathogenic bacterium antigen, adopt reinforced immunological method immunity bird inlay of the present invention and the corresponding composite IgY of preparation again, prepared anti-vagina Candida albicans, bacillus ducreyi specificity composite IgY have the ideal broad spectrum, not only vagina Candida albicans, the bacillus ducreyi antigen to direct immunization has very strong restraining effect, the antibodies of being surveyed tire higher (1: 512); Other two kinds of gynecological infection pathogenic bacterium that do not have direct immunization are had certain restraining effect too, and detected antibodies is tired and is still belonged to ideal.
Experimental example 3: the hen that experimental example 1 and two kinds of different composite antigens in the experimental example 2 are immune respectively produces the egg balanced mix, prepares anti-gynecological infection specificity composite IgY.Use " ELISA method " (enzyme linked immunosorbent assay) to use antigen with vagina Gardner bacillus, Trichomonas vaginalis, vagina Candida albicans, bacillus ducreyi as detecting respectively then, detect prepared anti-gynecological infection specific IgY these 4 kinds of antigenic antibodies of difference are tired.Detected result such as following table
????IgY | Detection antigen | Antibodies is tired |
Anti-gynecological infection specificity composite IgY | Vagina Gardner bacillus | ????1∶512 |
Trichomonas vaginalis | ????1∶512 | |
The vagina Candida albicans | ????1∶512 | |
Bacillus ducreyi | ????1∶512 |
Can see from above result, with the special complex antigen that contain multiple thalline composition of method of the present invention with the various pathogens preparation, use this special complex antigen again, use reinforced immunological method immunity bird inlay of the present invention, prepared anti-gynecological infection specificity composite IgY, gynecological infection pathogenic bacterium as the antigen direct immunization are had very high antibodies tire, reach suitable ideal effect.
Experimental example 4: the gynecology colpitis patient uses with " anti-gynecological infection specificity composite IgY " of the present invention clinical observation as the combination propellant treatment of raw material making
(1) in the patient who suffers from gynecology colpitis, picks out at random 20;
(2) " the anti-gynecological infection combination propellant " that adopts the present invention to prepare after the soup bottle is opened, poured soup in the PARI type compression atomizing air pump into, is sprayed directly on to the most concentrated positions of pathogenic agent such as vulva, vagina, uterine cervix, sprays secondary every day.
(3) every day the viewing test patient the symptom variation situation.
(4) after 1 week, 15 patient's inflammation disappear substantially with " anti-gynecological infection combination propellant " through spray, and 5 patient's inflammation alleviate.
(5) continuously spray with after 2 weeks, 19 patients' transference cure wherein, other also obviously alleviates 1 patient's symptom.
Embodiment 1: prepare anti-gynecological infection specificity composite IgY
Respectively according to the cultural characters and the optimal culture conditions of vagina Gardner bacillus, Trichomonas vaginalis, vagina Candida albicans, bacillus ducreyi, select the suitable culture base, carry out the strain of bacterium worm and breed in a large number, collect culture, purification, bacterial strain counting, dilution packing; Get vagina Gardner bacillus 10-20 hundred million, Trichomonas vaginalis purifying antigen protein 10 0ug makes 0.5ml bacterium liquid, adds the abundant mixing of 0.5ml freund adjuvant, makes the 1st kind of complex antigen; Get vagina Candida albicans 10-20 hundred million and bacillus ducreyi 10-20 hundred million and make 0.5ml bacterium liquid, add the abundant mixing of 0.5ml freund adjuvant, make the 2nd kind of complex antigen;
Respectively healthy anosis 200 of the U.S. Luo Man chickens of choosing of laying eggs are carried out immunity with prepared two kinds of complex antigens, every chest muscle injection 1ml, carry out the injection second time with same dosage after 7 days, injected for the third time with same dosage in 7 days at interval again, began to choose in the 20th day from the 1st injection back and get immune egg, stopped choosing egg to 10th month;
With synantigen not respectively the hen of immunity produce the egg balanced mix, with 0.1% potassium permanganate soaking disinfection 30 minutes, wash with sterile distilled water then, dry, place the broken eggshell of beating machiae, remove egg white with the yolk sieving, leaving and taking 6 times of distilled water of yolk adding stirs evenly, regulate PH to 5-8 with 1mol/l NaOH solution, 4 ℃ of following standing over night, with centrifugal 20 minutes of diluent 2000r/min, get supernatant with 15 times of molecular weight cut-off 200,000 ultra-filtration membrane ultrafiltration and concentration, concentrated solution is dried with freezing spray dry spray, promptly get the specificity composite IgY crude product, purifying crude is made the pure product of composite IgY by our company's patented technology (application number 00101270.3CN1307061A) method.
The preparation of embodiment 2, anti-gynecological infection specificity composite IgY washing lotion
Get anti-gynecological infection specificity composite IgY lyophilized powder 200g, allocate 200 liters of anti-gynecological infection specificity composite IgY solution by following prescription:
Anti-gynecological infection specificity composite IgY lyophilized powder 0.1%
Glycerine 2%
Eucalyptus oil 0.1%
Peppermint essence 0.2%
Borneol 0.1%
Ethanol 0.75%
Distilled water adds to 200 liters
Get 180 liters of distilled water earlier, add glycerine, composite IgY stirring and dissolving then respectively; Eucalyptus oil, peppermint essence, the borneol of getting formula ratio again add the ethanol stirring and dissolving of formula ratio, and under agitation slowly add in the aforementioned solution; Stir, measure the pH value of solution value, add distilled water to 200 liter at last, filter promptly between the 6.5-7.5.
The preparation of embodiment 3 anti-gynecological infection specificity composite IgY sprayss
Get anti-gynecological infection specificity composite IgY lyophilized powder 100g, allocate 200 liters of anti-gynecological infection specificity composite IgY solution by following prescription:
Anti-gynecological infection specificity composite IgY 0.05%
Peppermint essence 0.2%
Menthol 0.1%
Glycerine 2%
1% bright clear water solution is an amount of
Distilled water adds to 200 liters in right amount
Get 180 liters of distilled water earlier, add glycerine, composite IgY stirring and dissolving then respectively; Peppermint essence, menthol are dissolved in an amount of ethanol, under agitation slowly add in the aforementioned solution; Stir, measuring the pH value of solution value should be between 6.5-7.5; Add distilled water to 200 liter at last, with 1% bright clear water solution furnishing light green, filter the back can in the spray bottle promptly.
The preparation of embodiment 4 anti-gynecological infection specificity composite IgY effervescent tablets (capsule)
Anti-gynecological infection specificity composite IgY 0.5%
Citric Acid, usp, Anhydrous Powder 15%
Sodium bicarbonate 17%
Boric acid 5%
Lactose 31.1%
Microcrystalline Cellulose 30%
Suds-stabilizing agent 0.4%
The 10%PVP anhydrous alcohol solution is an amount of
Magnesium Stearate 1%
Except that IgY, boric acid, Magnesium Stearate, routinely with effervescent and other auxiliary materials and mixing,, add composite IgY and Magnesium Stearate, boric acid mixing then with the granulation of PVP anhydrous alcohol solution, dry, whole grain, compressing tablet or filled capsules are for the vagina external application.
The preparation of embodiment 5 anti-gynecological infection specificity composite IgY gelifying agents
Anti-gynecological infection specificity composite IgY 5%
Ka Bopuer-934 (Carbopol 934) 1%
HPMC (HPMC) 0.5%
Three ethanol eucalyptus 1%
Glycerine 10%
Ethanol 7.5%
Eucalyptus oil 1%
Peppermint essence 1%
Menthol 0.1%
Distilled water adds to 100%
Get menthol and add peppermint essence, eucalyptus oil dissolving mixing, add ethanol, the abundant mixing of glycerine, adding Ka Bopuer-934 and HPMC more respectively is uniformly dispersed, under agitation adding surplus distilled water mixes, add the IgY dissolving again, add three ethanol eucalyptus at last, reconcile between pH value 6.5-7.5, promptly make gelifying agent.
The preparation of embodiment 6 anti-gynecological infection specificity composite IgY ointments
Anti-gynecological infection specificity composite IgY 2%
Octadecanoic acid 6.1%
Stearyl alcohol 6.1%
Whiteruss 9.0%
White vaseline 6.1%
Span-80????????????????????????????0.85%
Glycerine 10%
Potassium sorbate 0.1%
Tween-80 4.5%
Menthol 0.2%
Distilled water adds to 100%
Oil water phase is heated to 80 ℃ respectively, and water is under agitation slowly added in the oil phase, stirs to be condensed to about 40 ℃, adds composite IgY solution, fully grinds to stir and makes the oil-in-water emulsifiable paste, promptly.
The preparation of embodiment 7 anti-gynecological infection specificity composite IgY vaginal suppositorys
Composite IgY 3g 1.5%
Gelatin 65.7g 32.85%
Glycerine 65.5g 32.75%
Distilled water (pure) 65.8g 32.90%
Full dose 200g 100%
Gelatin adding distil water swelling, the dissolving of heating, the glycerol adding mixing is chilled to below 40 ℃, and the composite IgY solution mixing reverse mould that adds with a small amount of dissolved in distilled water cools off promptly.
Claims (12)
1, a kind of anti-gynecological infection specific IgY is characterized in that the preparation method of this anti-gynecological infection specific IgY comprises the following steps:
Preferably cause the main pathogenic bacterium of gynecological infection, the main pathogenic bacterium antigen of preparation gynecological infection; Prepare immune egg; Prepare the main pathogenic bacterium specific IgY of anti-gynecological infection crude product; The purifying of the main pathogenic bacterium specific IgY of anti-gynecological infection crude product; Prepare anti-gynecological infection specific IgY.
2, anti-gynecological infection specific IgY as claimed in claim 1 is characterized in that the preparation method of this anti-gynecological infection specific IgY comprises the following steps:
Screening causes the germ of gynecological infectious diseases, on the basis of screening and the various gynecological infection pathogenic bacterium of sort merge, prepare single respectively or compound gynecological infection pathogenic bacterium antigen: according to cultural characters and the optimal culture conditions of various gynecological infection pathogenic bacterium, select the suitable culture base respectively, carrying out the strain of bacterium worm breeds in a large number, collect culture, purification, bacterial strain counting, dilution packing; With various single or blended gynecological infection pathogenic bacterium adding freund adjuvants, put into the high-speed homogenization machine again, the high speed pulverization homogenize forms milk-like slurry, makes single or compound gynecological infection pathogenic bacterium antigen.
Prepare immune egg: select to have (SPF) healthy bird inlay that the healthy no-special pathogen that facing out of high immunne response ability produces or laying eggs carries, every at chest muscle or subcutaneously inject prepared single or compound gynecological infection pathogenic bacterium antigen respectively, the interval is 7 days after injection for the first time, again for the second time with same dosage, method injection, injected the 3rd time with same dose in 7 days at interval again after the injection for the second time, the injection back was chosen since the 20th day and is got high immune egg for the first time, and corresponding different antigen, the immune egg of being searched is carried out coded markings.
The preparation of anti-gynecological infection pathogenic bacterium specificity composite IgY crude product: the immune respectively hen of synantigen does not produce the egg balanced mix, with 0.5% benzalkonium bromide solution or 0.1%KMnO4 solution or other similar thimerosals immune egg is soaked sterilization in 15-30 minute, the sterile distilled water flushing, dry, place beating machiae that eggshell is smashed, stir except that egg white stays 4-8 times of distilled water diluting of yolk adding with the yolk sieving, regulate 4-6 ℃ of following standing over night of PH to 5.5-6.0 with 1mol/l NaOH liquid or 1mol/l HCL liquid, diluent 1000-3000r/min high speed centrifugation 20 minutes, get supernatant liquor and hold back ultrafiltration and concentration 10-20 doubly with 170,000 molecular weight to 20, ten thousand molecular weight, concentrated solution is done promptly to get with the spray of cryospray desiccating method and is resisted gynecological infection specificity composite IgY crude product.
The preparation of the pure product of anti-gynecological infection pathogenic bacterium specificity composite IgY: the anti-gynecological infection pathogenic bacterium specific IgY crude extract that above technology is produced is crossed ion exchange column and gel exchange column respectively and is carried out purifying, promptly gets pure composite IgY;
3, anti-gynecological infection specific IgY as claimed in claim 1 or 2 is characterized in that causing that the pathogenic bacterium of gynecological infection comprise vagina Gardner bacillus, Trichomonas vaginalis, vagina Candida albicans, bacillus ducreyi among the preparation method of this anti-gynecological infection specific IgY.
4, anti-gynecological infection specific IgY as claimed in claim 1 or 2 is characterized in that this anti-gynecological infection specific IgY can make clinical acceptable forms.
5, anti-gynecological infection specific IgY as claimed in claim 2, the preparation that it is characterized in that gynecological infection pathogenic bacterium complex antigen described in the preparation method of this anti-gynecological infection specific IgY is such: at first cultivate preferred one or more pathogenic bacterium by the described method of claim 2, when selecting a kind of pathogenic bacterium, directly add freund adjuvant and make single antigen; When selecting various pathogens, then elder generation in 1-10: 1-10 or 1-10: 1-10: 1-10 or 1-10: 1-10: 1-10: 1-10 or 1-10: 1-10: 1-10: 1-10: the 1-10 ratio uniform mixing that the rest may be inferred, adds cultured more than one gynecological infection pathogenic bacterium then freund adjuvant and makes complex antigen.
6, anti-vagina Gardner bacillus, Trichomonas vaginalis, vagina Candida albicans, bacillus ducreyi various pathogens specificity composite IgY is characterized in that the preparation method of this anti-various pathogens specificity composite IgY comprises the following steps:
Respectively according to the cultural characters and the optimal culture conditions of vagina Gardner bacillus, Trichomonas vaginalis, vagina Candida albicans, bacillus ducreyi, select the suitable culture base, carry out the strain of bacterium worm and breed in a large number, collect culture, purification, bacterial strain counting, dilution packing; Get vagina Gardner bacillus 10-20 hundred million, Trichomonas vaginalis purifying antigen protein 10 0ug makes 0.5ml bacterium liquid, adds the abundant mixing of 0.5ml freund adjuvant, makes the 1st kind of complex antigen; Get vagina Candida albicans 10-20 hundred million and bacillus ducreyi 10-20 hundred million and make 0.5ml bacterium liquid, add the abundant mixing of 0.5ml freund adjuvant, make the 2nd kind of complex antigen;
Respectively healthy anosis 200 of the U.S. Luo Man chickens of choosing of laying eggs are carried out immunity with prepared two kinds of complex antigens, every chest muscle injection 1ml, carry out the injection second time with same dosage after 7 days, injected for the third time with same dosage in 7 days at interval again, began to choose in the 20th day from the 1st injection back and get immune egg, stopped choosing egg to 10th month;
With synantigen not respectively the hen of immunity produce the egg balanced mix, with 0.1% potassium permanganate soaking disinfection 30 minutes, wash with sterile distilled water then, dry, place the broken eggshell of beating machiae, remove egg white with the yolk sieving, leaving and taking 6 times of distilled water of yolk adding stirs evenly, regulate PH to 5-8 with 1mol/l NaOH solution, 4 ℃ of following standing over night, with centrifugal 20 minutes of diluent 2000r/min, get 15 times of molecular weight cut-off 200,000 ultra-filtration membrane ultrafiltration and concentration of supernatant, concentrated solution is done with freezing spray dry spray, promptly gets the specificity composite IgY crude product, with above IgY dissolving crude product, earlier after DEAE-SephadexA50 column chromatography and SephadexG200 gel filtration chromatography,, promptly get and resist vagina Gardner bacillus purifying crude, Trichomonas vaginalis, the vagina Candida albicans, bacillus ducreyi various pathogens specificity composite IgY.
7, a kind of preparation method of anti-gynecological infection washing lotion is characterized in that this method is:
Get anti-gynecological infection specificity composite IgY lyophilized powder 200g, allocate 200 liters of anti-gynecological infection specificity composite IgY solution by following prescription:
Anti-gynecological infection specificity composite IgY lyophilized powder 0.1%
Glycerine 2%
Eucalyptus oil 0.1%
Peppermint essence 0.2%
Borneol 0.1%
Ethanol 0.75%
Distilled water adds to 200 liters
Get 180 liters of distilled water earlier, add glycerine, composite IgY stirring and dissolving then respectively; Eucalyptus oil, peppermint essence, the borneol of getting formula ratio again add the ethanol stirring and dissolving of formula ratio, and under agitation slowly add in the aforementioned solution; Stir, measure the pH value of solution value, add distilled water to 200 liter at last, filter promptly between the 6.5-7.5.
8, a kind of anti-gynecological infection spray preparing process is characterized in that this method is:
Get anti-gynecological infection specificity composite IgY lyophilized powder 100g, allocate 200 liters of anti-gynecological infection specificity composite IgY solution by following prescription:
Anti-gynecological infection specificity composite IgY 0.05%
Peppermint essence 0.2%
Menthol 0.1%
Glycerine 2%
1% bright clear water solution is an amount of
Distilled water adds to 200 liters in right amount
Get 180 liters of distilled water earlier, add glycerine, composite IgY stirring and dissolving then respectively; Peppermint essence, menthol are dissolved in an amount of ethanol, under agitation slowly add in the aforementioned solution; Stir, measuring the pH value of solution value should be between 6.5-7.5; Add distilled water to 200 liter at last, with 1% bright clear water solution furnishing light green, filter the back can in the spray bottle promptly.
9, the preparation method of a kind of anti-gynecological infection effervescent tablet (capsule) is characterized in that this method is:
Anti-gynecological infection specificity composite IgY 0.5%
Citric Acid, usp, Anhydrous Powder 15%
Sodium bicarbonate 17%
Boric acid 5%
Lactose 31.1%
Microcrystalline Cellulose 30%
Suds-stabilizing agent 0.4%
The 10%PVP anhydrous alcohol solution is an amount of
Magnesium Stearate 1%
Except that IgY, boric acid, Magnesium Stearate, routinely with effervescent and other auxiliary materials and mixing,, add composite IgY and Magnesium Stearate, boric acid mixing then with the granulation of PVP anhydrous alcohol solution, dry, whole grain, compressing tablet or filled capsules are for the vagina external application.
10, a kind of preparation method of anti-gynecological infection gelifying agent is characterized in that this method is:
Anti-gynecological infection specificity composite IgY 5%
Ka Bopuer-934 (Carbopol 934) 1%
HPMC (HPMC) 0.5%
Three ethanol eucalyptus 1%
Glycerine 10%
Ethanol 7.5%
Eucalyptus oil 1%
Peppermint essence 1%
Menthol 0.1%
Distilled water adds to 100%
Get menthol and add peppermint essence, eucalyptus oil dissolving mixing, add ethanol, the abundant mixing of glycerine, adding Ka Bopuer-934 and HPMC more respectively is uniformly dispersed, under agitation adding surplus distilled water mixes, add the IgY dissolving again, add three ethanol eucalyptus at last, reconcile between pH value 6.5-7.5, promptly make gelifying agent.
11, a kind of preparation method of anti-gynecological infection ointment is characterized in that this method is:
Anti-gynecological infection specificity composite IgY 2%
Octadecanoic acid 6.1%
Stearyl alcohol 6.1%
Whiteruss 9.0%
White vaseline 6.1%
Span-80??????????????????????0.85%
Glycerine 10%
Potassium sorbate 0.1%
Tween-80 4.5%
Menthol 0.2%
Distilled water adds to 100%
Oil water phase is heated to 80 ℃ respectively, and water is under agitation slowly added in the oil phase, stirs to be condensed to about 40 ℃, adds composite IgY solution, fully grinds to stir and makes the oil-in-water emulsifiable paste, promptly.
12, a kind of preparation method of anti-gynecological infection vaginal suppository is characterized in that this method is:
Composite IgY 3g 1.5%
Gelatin 65.7g 32.85%
Glycerine 65.5g 32.75%
Distilled water (pure) 65.8g 32.90%
Full dose 200g 100%
Gelatin adding distil water swelling, the dissolving of heating, the glycerol adding mixing is chilled to below 40 ℃, and the composite IgY solution mixing reverse mould that adds with a small amount of dissolved in distilled water cools off promptly.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 03136958 CN1454901A (en) | 2003-06-10 | 2003-06-10 | Gynaecological anti-infective specificity IgY and its combined preparation |
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