LU101793B1 - Single-chain antibody TRsAb2 against ToRSV and preparation method thereof - Google Patents
Single-chain antibody TRsAb2 against ToRSV and preparation method thereof Download PDFInfo
- Publication number
- LU101793B1 LU101793B1 LU101793A LU101793A LU101793B1 LU 101793 B1 LU101793 B1 LU 101793B1 LU 101793 A LU101793 A LU 101793A LU 101793 A LU101793 A LU 101793A LU 101793 B1 LU101793 B1 LU 101793B1
- Authority
- LU
- Luxembourg
- Prior art keywords
- trsab2
- chain antibody
- torsv
- sequence
- variable region
- Prior art date
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- C—CHEMISTRY; METALLURGY
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- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
- C07K2317/622—Single chain antibody (scFv)
Landscapes
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- Virology (AREA)
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- Peptides Or Proteins (AREA)
Claims (2)
- | | LU101793 | Revendications | 1. Un anticorps à chaîne unique TRsAb2 contre ToRSV, ayant une séquence d'acides ) aminés comme indiqué dans SEQ ID NO.1.| 2. Anticorps à chaîne unique TRsAb2 selon la revendication 1, consistant en une région | de marqueur histidine 15aa, une région variable de chaîne lourde ToRSV 184aa, un | peptide de liaison 15aa et une région variable de chaîne légère ToRSV 102aa liée en | liaison inverse, connectées en séquence, d'une longueur de 316 résidus d'acides : aminés, où les séquences de la région variable de chaîne lourde 184aa et de la ; région. variable de chaîne légère 102aa sont dérivées de la séquence d'un anticorps | monoclonal produit par la souche cellulaire d'hybridome CGMCC18320.| 3. Anticorps à chaîne unique TRsAb2 selon la revendication 1, dans lequel la séquence | du gène codant de l'anticorps monocaténaire TRsAb2 est telle que représentée dans | SEQ ID NO.2.| 4. Anticorps à chaîne unique TRsAb2 selon la revendication 3, dans lequel la séquence | du gène codant de l'anticorps monocaténaire TRsAb2 a une longueur de 948 pb, | dans laquelle la séquence du gène codant de la région d'étiquette histidine a une | longueur de 45 pb, le codage la séquence du gène de la région variable de la chaîne ; lourde ToRSV 184aa a une longueur de 552 pb, la séquence du gène codant du | peptide de liaison est de 45 pb et la séquence du gène codant de la région variable | de la chaîne légère du ToRSV 102aa a une longueur de 306 pb.| 5. Anticorps à chaîne unique TRsAb2 selon la revendication 3, dans lequel l'amont de la | séquence du gène codant de l'anticorps monocaténaire TRsAb2 comprend en outre | des éléments d’expression : promoteurs Lac, opérons Lac et sites de liaison aux | ribosomes (RBS).| 6. Anticorps à chaîne unique TRsAb2 selon la revendication 4, dans lequel un vecteur | d'expression d'anticorps à chaîne unique TRsAb2 est PUC57, son site d'insertion est | un site de coupure de l'enzyme de restriction EcoRV et sa souche hôte d'expression | est une souche E. coli DH5a.' 7. Anticorps à chaîne unique TRsAb2 selon la revendication 1, dans lequel un procédé | de production de l'anticorps à chaîne unique TRsAb2 comprend les étapes suivantes: | (1) construction d'un vecteur d'expression d'anticorps a chaine unique TRsAb2: | insertion d'une séquence de gène représentée dans SEQ ID NO. 2 dans un site | EcoRV sur un vecteur PUC57 pour obtenir un vecteur d'expression d'anticorps à | chaîne unique TRsAb2; | (2) introduire le vecteur d'expression d'anticorps à chaîne unique TRsAb2 dans E. coli | DH5a pour obtenir une souche d'expression d'anticorps monocaténaire TRSAB2; | (3) expression auto-induite de l'anticorps à chaîne unique TRsAb2: inoculation de la | souche d'expression d'anticorps à chaîne unique TRSAB2 dans un milieu | d'expression auto-inducteur, et fermentation à une température de 16-40°C pendant | 12-24 heures pour obtenir un bouillon de fermentation du produit, dans lequel un litre | du milieu d'expression d'auto-induction comprend 1,0-5,0 g d'extrait de levure, 5,0- 10,0 g de tryptone, 5,0-15,0 g de poudre de maïs, 0,1-1,5 g de saccharose, 4,0-10,0 | g de phosphate dipotassique, 3,5-9,0 g de dihydrogénophosphate de potassium, 2,0- | 6,0 g d'hydrogénophosphate de diammonium, 0,2-1,6 g de sulfate de magnésium,| 2,0-5,0 mg de chlorure de calcium, 0,5-3,5 mg de chlorure de cobalt, 0,5-1,5 mg de 0101798 chlorure de cuivre, 1,2-5,0 mg de sulfate de manganèse, 3,2-8,0 mg de molybdate desodium, 0,1-0,5 mg d'acide borique, 1,0-8,0 mg de chlorure ferrique, 0,5-3,0 mg de| sulfate de zinc, 1,0-10,0 g de du glycérol et 20 à 300 mg d'isopropylthio-B-D-| galactoside (IPTG);| (4) séparation et purification de l'anticorps à chaîne unique TRsAb2: centrifugation| du bouillon de fermentation obtenu à l'étape (3), collecte des cellules bactériennes,| remise en suspension dans une solution tampon phosphate, realisation d’une| ultrasonication, puis re-centrifugation, collecte du surnageant, charger le surnageant| sur une colonne d'ions nickel, éluer avec de imidazole selon une concentration en| gradient, collecter l'éluat d'anticorps TRsAb2 à chaîne unique et effectuer la dialyse et| la lyophilisation pour obtenir l'anticorps TRsAb2 à chaîne unique.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| LU101793A LU101793B1 (en) | 2020-05-15 | 2020-05-15 | Single-chain antibody TRsAb2 against ToRSV and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| LU101793A LU101793B1 (en) | 2020-05-15 | 2020-05-15 | Single-chain antibody TRsAb2 against ToRSV and preparation method thereof |
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| Publication Number | Publication Date |
|---|---|
| LU101793B1 true LU101793B1 (en) | 2020-09-21 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| LU101793A LU101793B1 (en) | 2020-05-15 | 2020-05-15 | Single-chain antibody TRsAb2 against ToRSV and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| LU (1) | LU101793B1 (fr) |
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2020
- 2020-05-15 LU LU101793A patent/LU101793B1/en active IP Right Grant
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