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How to Improve the Fusion Rate of Monoclonal Antibodies
<p><b>Abstract</b>:&nbsp; &nbsp;At the beginning of performing monoclonal antibody test, the failure of cell fusion or low fusion rate happens sometimes. Now, some factors for influencing fusion rate are summarized, which may be helpful for experimenters.<br><br><b>Keywords</b>: Monoclonal Antibodies, Fusion Rate, Antibodies Fusion</p><p><br></p><p><b>1. Blood Titer Test</b><br>Before the fusion, the mouse blood should be collected for testing the titer. The titer higher than 20,000 is suitable for the fusion. The fusion effect may not be satisfactory if the titer is lower than 20,000.<br></p><p><br></p><p><b>2. Cell State</b><br>The state of myeloma cells SP2/0 is very important and should be kept without any pollution and large scale death. The logarithmic growth phase of cells should be maintained before the fusion.<br></p><p><br></p><p><b>3. PEG and Fatal Bovine Serum</b><br>The fusion rate depends on the quality of PEG and fatal bovine serum which should be subject to the pilot experiment screening between different brands and batch numbers. Selecting the best quality PEG and fatal bovine serum can ensure the successful rate of fusion efficiently.<br></p><p><br></p><p><b>4. Cell Ratio</b><br>During the fusion, the ratio of SP2/0 and splenocytes should be noticed as well. Although 10:1-1:10 is applicable for the successful fusion, the best fusion efficiency is between 2:1-1:2. No matter which ratio, the two kinds of cells don't contain serum components and should be mixed well. During the fusion process, the fusion tube should be always rotated. Dropping PEG and terminating the fusion should be slowly first and fast afterwards to avoid the great impact on the cell.<br></p><p><br></p><p><b>5. Feeder Layer Cells</b><br>Feeder layer cells can also influence the fusion effect. Feeder cells of hybridoma can be chosen from mouse peritoneal macrophages, splenocytes and thymocytes. The peritoneal macrophages is the best choice. Meanwhile, the amount of feeder cells in each well should be also noticed. Excessive cells will fight for medium nutrition, making the growth of hybridoma cells difficult to maintain. Deficient cells can't serve as the feeder layer. Hence, only the reasonable cell amount can achieve the good effect.<br></p><p><br></p><p><b>6. Medium Additives</b><br>The intelligent use of some additives has a good prospect. E.g. the hybridoma cell culture medium additive developed by FineTest can efficiently extend the service life of medium, improve the growth speed of hybridoma cells and increase fusion rate. Under unsatisfactory experimental conditions, the fusion rate can be still maintained well.<br></p><p><br></p><p><img src=""><br></p><p><b>Figure 1.</b>&nbsp; Hybridoma Cell Culture Medium Additive(100X)<br></p><p><br></p><p><img src=""><br></p><p><b>Figure 2.</b> Comparison between fusion cells without medium additives and hybridoma cells with medium additives.<br></p><p><br></p><p><b>7. Conclusion</b><br>There are other factors for influencing fusion rate. E.g.: the number of cells in each well after the fusion, centrifugal rotational speed and time, the strength for pipetting cells etc. During the actual experimental process, it's important to accumulate more experience.<br></p>
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How to Improve the Fusion Rate of Monoclonal Antibodies00