Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA.
BACKGROUND
A simple filter paper method was developed for, the transport and storage of monoclonal antibodies (Mabs) at room temperature or -20 degrees C after spotting on filter paper, for subsequent serotyping of outer membrane antigens of N.meningitidis by dot-blot ELISA.


METHODS
Monoclonal antibodies (Mabs) were spotted within a 0.5-1 cm diameter area of Whatman grade 903 paper, which were stored individually at room temperature or at -20 degrees C. These MAbs were stored and analyzed after periods of one week, 4 weeks, 12 months, or 13 years in the case of frozen Mab aliquots, or after 4 weeks at -20 degrees C or at room temperature (RT) in the case of Mabs dried on filter paper strips. Assays were performed in parallel using dot-blot ELISA. In addition to the MAbs specific for serotyping class 1, 2 or 3, we used a larger number of Mabs for polysaccharides, lipooligosaccharides (LOS), class 5 and cross-reactive antigens for native outer membrane of N.meningitidis. The Mabs dried on filter paper were eluted with phosphate-buffered saline (PBS) containing 0.2% gelatin.


RESULTS
Mabs of the isotypes IgG and IgM dried on filter papers were not affected by duration of storage. The detection by serotyping Mabs was generally consistent for dried filter paper MAb samples stored frozen for over 1 year at -20 degrees C, and although decreased reactive antibody titers were found after storage, this did not interfere with the specificity of the Mabs used after 13 years as dry spots on filter paper.


CONCLUSIONS
The use of filter paper is an inexpensive and convenient method for collecting, storing, and transporting Mab samples for serotyping studies. In addition, the samples occupy little space and can be readily transported without freezing. The efficiency of using immunoglobulin G (IgG) or M (IgM) eluted was found to be consistent with measurement of IgG or IgM titers in most corresponding, ascites Mabs stored frozen for over 1 year. The application of meningococcal typing methods and designations depend on the question being asked.
Ferraz
Aline S
AS
Immunology Section, Adolfo Lutz Institute, São Paulo, São Paulo, Brazil. alineseneme@hotmail.com
Belo
Elza F T
EF
Coutinho
Ligia M C C
LM
Oliveira
Ana P
AP
Carmo
Andréia M S
AM
Franco
Daniele L
DL
Ferreira
Tatiane
T
Yto
André Y
AY
Machado
Marta S F
MS
Scola
Monica C G
MC
De Gaspari
Elizabeth
E
D000818:Animals; D000911:Antibodies, Monoclonal; D015373:Bacterial Typing Techniques; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005374:Filtration / Q000295:instrumentation; D006801:Humans; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008851:Micropore Filters; D009345:Neisseria meningitidis / Q000145:classification* / Q000276:immunology; D010209:Paper; D012703:Serotyping / Q000379:methods; D013048:Specimen Handling / Q000379:methods*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007075:Immunoglobulin M