Characterizing antibody cross-reactivity for immunoaffinity purification
of analytes prior to multiplexed liquid chromatography-tandem mass spectrometry.
Characterization of SF-Derived Exosomes Isolated by the Immunoaffinity Purification
In particular, the tag appended to the N-terminus of the fusion protein is necessary for the immunoaffinity purification
with M1 monoclonal antibody, while M2 is position-insensitive.
The material covers basic methods of antigen selection and preparation, antibody production, and reagent manufacture, as well as a number of advanced protocols such as techniques for making new monoclonal cell lines, producing monoclonal antibodies, immunocapture PCR, immunoaffinity purification
, and immunoelectron microscopy in plant virus diagnosis.
Heavy-isotope-labeled ([sup.13]C-[sup.15]N-Lys/Arg) recombinant tau was spiked into CSF samples before immunoaffinity purification
and subsequent digestion and analysis.
The 2 methods were relatively poorly correlated, which may be due to the improved specificity of the immunoaffinity purification
approach (see Fig.
The immunoaffinity purification
of the sample therefore yielded a much cleaner preparation than the SPE procedures, which result in multiple peaks even when analyzed by tandem MS (10,11).
Fractions containing proteins smaller than 20 kDa were collected and subjected to immunoaffinity purification
as described above.
In the case of the reports in this issue of Clinical Chemistry (1, 2), the samples were prepared for analysis by a single-step immunoaffinity purification
that can be carried out in large numbers using a standard 96-well microtiter plate format.
of creatine kinase-MB from human, dog, and rabbit heart with use of a monoclonal antibody specific for CK-MB.
and gas chromatography-mass spectrometric quantitation of 3-alkyladenines in urine: metabolism studies and basal excretion levels in man.