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We found that our method of microvesicle isolation identified a group of proteins that were not detected among the most abundant proteins in routinely processed sera (Venn diagram, Fig. 1E).
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From this point onwards the sera were processed as chicken sera.
Reactions were processed using Sera-Mag™ magnetic carboxylate-modified microparticles (Seradyn™, Indianapolis, IN, USA) to remove excess fluorescent terminators before loading onto GE Healthcare MegaBACE 4000 capillary DNA sequencers.
This makes it possible to process many sera at the same time, using multichannel pipettes.
These sera were randomly distributed and were few as compared to the large number of sera processed in the study.
Blood samples were processed promptly and sera were stored at -20°C or colder pending testing at the national reference laboratory.
Pre-immune (week 0) and pre-challenge (week 5) tail blood samples of individual mice were collected, processed to obtain sera and diluted 1 100 in PBS.
Study design: From a total of 190 human serum samples from 43 healthy children, 24 pregnant women and 24 patients with bone marrow transplants, 111 sera were processed by indirect immunofluorescence assay for the detection of IgG1, IgG2, IgG3 and IgG4 specific antibodies.
Sera were processed by HAI and MNA.
Sera were processed in replicates and deposited onto the MALDI plate in duplicates.
When immunized sera were processed for micro-virus neutralization assays using 100 TCID50 of CA/04/09 in Madin-Darby canine kidney (MDCK) cells, administration of the reverse-genetics vaccine demonstrated efficient virus neutralizing activity which correlated comparably well with the vaccine-induced HI responses (data not shown).
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