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兔单抗亲和力和灵敏度更高
相对于鼠单抗（解离常数Kd在10-9-10-10M水平），兔单抗的亲和力可提高100-1000倍（Kd在10-10-10-12M水平）。实际使用时，兔单抗的工作浓度更低，灵敏度更高，因此产生的背景更低，大大降低了假阳性的发生。用作免疫组化等用途时，有时甚至可免去抗原修复的步骤。
兔单抗能识别更多表位
很多蛋白在人类和啮齿动物中同源性很高，这些保守表位在小鼠体内免疫原性很弱，不容易产生高质量抗体。而兔做为宿主，可以更好地识别这些表位，产生针对更多表位的抗体。当然，兔也适合生产针对鼠类蛋白的抗体。
Fusion and Cloning
Add 1 ml of PEG solution to the pellet over a 1 minute period, continually stirring the cells.
Continue stirring for an additional 1 minutes.
Stop the fusion by adding Medium B while constantly stirring.
1 ml over 1 minute
3 ml over 1 minute
10 ml over 1 minute
Incubate for 5 minutes in a water bath at 37oC.
Slowly add 40 ml of Medium A.
Centrifuge the cells at 400 g for 7 minutes.
Discard the supernatant and wash the cell in 40 ml of Medium A.
Slowly resuspend the pellet in 10 ml of Medium C.
Transfer to a T75 flask containing 40 ml of Medium C.
Incubate 16-24 hours at 37oC, 5% CO2.
Thaw Medium D and mix.
Transfer the cells from the flask into 2x50 ml centrifuge tubes and centrifuge at 400 g for 10 minutes.
Discard the supernatants and tap to loosen the pellets.
Combine the pellets and transfer the cells to Medium D. Mix gently by swirling the tube.
Let sit for 30 minutes at 37oC, 5% CO2.
Plate 9.5 ml of cells into 10-100 mm Petri dishes. Tilt the plates to level the mixture.
Transfer the plates to a Tupperware container containing a Petri dish with 10 ml sterile water.
Incubate plates at 37oC, 5% CO2.
Maintenance
After 10-14 days, examine the plates for the presence of colonies.
Remove isolated colonies from the plates using a pipettor with a sterile tips. Set the pipettor for 10 ml.
Pipet each colony into a separate well of 96-well plates contain 200 ml of Medium E.
Incubate the plates at 37oC, 5% CO2 for 1-4 days without feeding.
Remove the supernatants from the wells and test. Refeed the wells with 200 ml Medium E or other Hybridoma expansion medium.
Recloning in ClonaCell-HY
Note: More than 95% of the colonies will be monoclonal when selected by ClonaCell-HY. Recloning can be done to ensure stability.
Once the cells are growing well in 24-well plates, resuspend the cells with a 1 ml pipet.
Remove 10 ml of the cell suspension and add to 1.0 ml of Medium A. Mix well.
Remove 100 ml of this suspension and add to a tube containing 10 ml of Medium D. Mix well and add to a 100 mm Petri dish.
Spread evenly by tilting the plates. Incubate at 37oC, 5% CO2 as previously described.
Repeat for each clone.
After 10-14 days, select colonies and transfer to 96-well plates before testing.
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