Luminex® Single Antigens

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  • We perform 5 washes and adjust the vacuum pressure to 100-125mbars and hold each wash for 5 seconds. Thanks for your comments.
  • It makes perfect sense. How many washes do you do after the first incubation step when using the manifold? Could you off-set poor washing that you get with low vaccuum pressure simply by increasing the number of washes? In my experience, increasing the number of washes from 3 to 4 can certainly even out any differences you get due to poor "flicking" technique.
  • Perhaps it's all in how well technologists flick and wash out non-HLA, IgG antibodies. In our experience the pressure on the vacuum manifold has to be precise, otherwise we start loosing beads (perhaps by crushing them). If the pressure is low our washing is poor; if the pressure is too high we loose beads. I can imagine that technologists are taught to flick hard without fear for loosing beads? This is probably a good thing when it comes to washing and removing non-HLA antibodies that my neutralize the anti-human IgG conjugate.
  • Very interesting. Any ideas regarding possible factors that contribute to these differences?
  • Great! We do all of our testing using filter plates and in some cases have seen data that suggest differences in results based on which method (spin or filter plate) is used.
  • Hi Renato,

    Thanks for your comments. Yes, we just started validating the rapid protocol using filter plate washes and the preliminary results look good. In addition, two labs that participated in the multicenter ROB protocol evaluation study (late breaking abstract presented at ASHI) used filter plate washing technique. I can put you in touch with them if you are interested.
  • Rob, this was a nice study to show that some of these Solid phase assays can further be optimized without compromising sensitivity. I am looking forward to trying out this procedure in the laboratory. I wonder if you have tried the filter plate instead of the spin method?
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