Natural alpha-syn oligomer antibody preparation method and product thereof, and applications of natural alpha-syn oligomer antibody
An oligomer and antibody technology, applied in the field of preparation of natural α-syn oligomer antibody, can solve the problems of complex preparation process and difficult operation, etc.
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Embodiment 1
[0107] This embodiment provides a kind of NAbs-α-syn, its preparation method comprises the following steps:
[0108] Add polyacrylamide gel beads to pH=10 buffer solution containing 0.6M sodium citrate and 0.2M sodium carbonate to wet the gel beads; Combined α-syn oligomers, mix well, shake for 1 h, centrifuge at 1200g for 1 min, discard the liquid phase; use pH=9.0 containing 3.0M NH 2 (CH 2 ) 2 Resuspend in OH buffer, shake for 2.5 hours, centrifuge at 1200g for 1min, discard the liquid phase; resuspend with PBS buffer, shake for 15min, centrifuge at 1200g for 1min, discard the liquid phase; use 1M NaCl solution Resuspend, shake for 15 minutes, centrifuge at 1200g for 1min, and discard the liquid phase; resuspend with PBS buffer, shake for 15 minutes, centrifuge at 1200g for 1min, discard the liquid phase, and repeat this process once to obtain the α -syn oligomer affinity gel; Dilute IVIG 10 times with PBS buffer and load it on α-syn oligomer affinity gel column, elute u...
Embodiment 2
[0110] This embodiment provides a kind of NAbs-α-syn, its preparation method comprises the following steps:
[0111] Add polyacrylamide gel beads to pH=10 buffer solution containing 0.6M sodium citrate and 0.2M sodium carbonate to wet the gel beads; Combined α-syn oligomers, mix well, shake for 0.5h, centrifuge at 1000g for 5min, discard the liquid phase; use pH=9.0 containing 3.0M NH 2 (CH 2 ) 2 Resuspend in OH buffer, shake for 1h, centrifuge at 1000g for 5min, discard the liquid phase; resuspend with PBS buffer, shake for 5min, centrifuge at 1000g for 5min, discard the liquid phase; use 1M NaCl solution Resuspend, shake for 5 minutes, centrifuge at a speed of 1000g for 5 minutes, and discard the liquid phase; resuspend with PBS buffer, shake for 5 minutes, centrifuge at a speed of 1000g for 5 minutes, and discard the liquid phase. This process is repeated 3 times to obtain the α- Syn oligomer affinity gel; IVIG was diluted 15 times with PBS buffer and loaded on the α-syn...
Embodiment 3
[0113] This embodiment provides a kind of NAbs-α-syn, its preparation method comprises the following steps:
[0114] Add the agarose gel to the buffer solution containing 0.6M sodium citrate and 0.2M sodium carbonate at pH=10 to wet the gel beads; add cross-linked α- Syn oligomer, mix well, shake for 2h, centrifuge at 3000g for 0.5min, discard the liquid phase; use pH=9.0 containing 3.0M NH 2 (CH 2 ) 2 Resuspend in OH buffer, shake for 5 hours, centrifuge at 3000g for 0.5min, discard the liquid phase; resuspend with PBS buffer, shake for 60min, centrifuge at 3000g for 0.5min, discard the liquid phase; use 1M NaCl The solution was resuspended, shaken for 60 minutes, centrifuged at a speed of 3000g for 0.5min, and the liquid phase was discarded; resuspended with PBS buffer, shaken for 60 minutes, centrifuged at a speed of 3000g for 0.5min, and the liquid phase was discarded. This process was repeated 5 times to obtain The α-syn oligomer affinity gel; the IVIG is diluted 20 ti...
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