1, the fusion rate is low, the number of positive holes is small
1 immune problem. Due to weak immunogenicity or improper immunization pathway, the immunity is weak and the titer is low.
2 temperature, time, PEG molecular weight, time of action, etc. during the fusion process.
3 Whether the spleen cells have taken out enough cells to interfere with tissue debris.
4 The pre-fusion myeloma cells are in good condition, and the cells are round, translucent and logarithmically grown.
2, the overall affinity of the monoclonal antibody is low
1 immune problem. Improper immune pathways or immunogenic problems.
2 The cloning cell screening process is missing, the screening method is improper or the detection method is improper, resulting in no fused to high affinity fusion cells.
3 Due to the inappropriate growth environment of the subcloned cells, the chromosomes of the fusion cells are lost, the secretion characteristics are changed, and the affinity changes from high to low. At this time, the culture solution needs to be changed in time. When the cells grow to a certain density, the medium begins to change color. It affects the growth of fused cells, causing chromosome loss; controlling the density of cells, the density is too large, so that the newly added medium will not be consumed quickly, affecting the growth of fused cells, causing chromosome loss; do not operate cells too frequently, when cells When the growth density is not very large, do not operate frequently, otherwise the cells are prone to death or the growth pattern changes significantly.
The operation is meticulous to prevent cells from being contaminated by microorganisms such as mycoplasma.
3, antibody affinity changes from high to low
1 The culture medium is not changed in time. When the cells grow to a certain density, the medium begins to change color, affecting the growth of the fused cells, causing chromosome loss or trait changes.
2 Control the density of the cells. If the density is too large, the newly added medium will not be consumed quickly, affecting the growth of the fused cells, causing chromosome loss or trait changes.
3 Operate the cells too frequently. When the cell growth density is not very large, do not operate frequently, otherwise the cells are prone to death or the growth pattern changes significantly.
4 cells are contaminated by microorganisms such as mycoplasma.
5 subcloning more than 5 times, resulting in a frequent change in the cell growth environment, weak positive hybridoma cells with chromosome loss or trait changes.
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