Crystals and structure of Synagis Fab
View Patent ↗The present invention provides crystalline forms and three-dimensional atomic structure coordinates of Synagis Fab. The present invention further provides methods of using such three-dimensional atomic structure coordinates for designing and identifying Synagis binding agents. The present invention also provides machine readable media embedded with the three-dimensional atomic structure coordinates of Synagis Fab and methods of using them.
1. A crystal comprising Synagis Fab in crystalline form, wherein Synagis Fab comprises a complex of Synagis light chain consisting of the sequence of SEQ ID NO:7 and N-terminal fragment of Synagis heavy chain consisting of the sequence of SEQ ID NO: 2, wherein the crystal has a space group of P2 1 2 1 2 1 and unit cell parameters of a=77.36±0.2 Å, b=103.92±0.2 Å and c=68.87±0.2 Å and is of diffraction quality.
2. The crystal of claim 1 , which is a native crystal.
3. The crystal of claim 1 , which is characterized by a diffraction pattern of FIG. 2 .
4. The crystal of claim 1 , which is produced by a method comprising the step of:
incubating a mixture in a closed container over a reservoir solution comprising a precipitant in a closed container under conditions suitable for crystallization until the crystal forms, wherein said mixture comprises a volume of a solution comprising Synagis Fab and a volume of the reservoir solution.
5. The crystal of claim 4 wherein the precipitant is PEG with an average molecular weight between 3350 and 8000.
6. The crystal of claim 4 wherein the precipitant is PEG with an average molecular weight of 4000.
7. The crystal of claim 4 , wherein the precipitant is present in a concentration between 14% and 20% (w/v).
8. The crystal of claim 4 wherein the precipitant is present in a concentration of 15%.
9. The crystal of claim 4 , wherein the solution further comprises between 50 mM Tris and 100 mM Tris.
10. The crystal of claim 4 , wherein the solution further comprises between 7% and 20% 2-propanol.
11. The crystal of claim 4 , wherein the solution further comprises 10% 2-propanol.
12. The crystal of claim 4 , wherein the solution has a pH of between 6.4 and 11.
13. The crystal of claim 4 , wherein the solution has a pH of 8.5.
14. The crystal of claim 4 , which is produced by incubating the mixture comprising Synagis Fab and reservoir solution at a temperature of between 4° C. and 27° C.
15. The crystal of claim 4 , which is produced by incubating the mixture comprising Synagis Fab and reservoir solution at a temperature of between 17° C. and 22° C.
16. The crystal of claim 4 , which is produced by incubating the mixture comprising Synagis Fab and reservoir solution at a temperature of 20° C.
17. A method of making the crystal of claim 1 , comprising:
(a) mixing a volume of a solution comprising a Synagis Fab polypeptide with a volume of a reservoir solution comprising a precipitant; and
(b) incubating the mixture obtained in step (a) over the reservoir solution in a closed container, under conditions suitable for crystallization until the crystal forms.
18. The method of claim 17 , wherein the precipitant is PEG with an average molecular weight between 3350 and 8000.
19. The method of claim 17 , wherein the precipitant is present in a concentration between 14% and 20% (w/v).
20. The method of claim 17 , wherein the solution further comprises 50 mM Tris.
21. The method of claim 17 , wherein the solution further comprises between 7% and 20% 2-propanol.
22. The method of claim 17 , wherein the solution has a pH of between 6.4 and 11.
23. The method of claim 17 , wherein the mixture comprising Synagis Fab and reservoir solution is incubated at a temperature of between 4° C. and 27° C.
24. The method of claim 17 , wherein the mixture comprising Synagis Fab and reservoir solution is incubated at a temperature of between 17° C. and 22° C.
25. The method of claim 17 , wherein the mixture comprising Synagis Fab and reservoir solution is incubated at a temperature of 20° C.