By Bernd Lepenies
This quantity summarizes present state-of-the-art equipment with regards to carbohydrate-based vaccines, from the identity of an appropriate carbohydrate antigen through the coaching of glycoconjugate vaccines to the characterization of vaccine applicants for his or her use in pre-clinical and medical reviews. Written within the hugely profitable Methods in Molecular Biology series layout, chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and key tips about troubleshooting and warding off recognized pitfalls.
Authoritative and useful, Carbohydrate-Based Vaccines: tools and Protocols goals to be a worthwhile handbook that would facilitate learn within the box of carbohydrate-based vaccines.
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Additional resources for Carbohydrate-Based Vaccines: Methods and Protocols
This will generate a template mask with circular boundaries. 2. 1 and adjust the GAL file so that the mask aligns with the actual spots on the slide. Inspect the whole slide and flag spots as “bad” that are either missing or contain defects in printing. These glycans will be excluded from further analysis. Save these modified settings as a GenePix Setting (GPS) file. 1. 6, and load the GPS file from the previous step. Finely adjust the size and position of the each circle mask so that it fits with individual spot size perfectly (see Note 13).
3. Scan the finished 384-well plates in FLx800 Microplate Fluorescence Reader for any missing samples. 4. Seal plates with aluminum seal to prevent evaporation and photobleaching of the dye, then cover with lids and store at 4 °C till use. 5. Spin down the source plate at 1000 rpm for 5 min before printing. 2 Set Up Robotic Microarray Printer 1. Program the printer so that each slide accommodates 16 arrays and each glycan is printed in duplicate (Fig. 2). 2. Sonicate printer pins for 15 min in Milli-Q water and blow-dry with argon.
36. and immobilization on epoxide-modified glass slides. Methods Mol Biol 808:155–165 Di Cristina M, Nunziangeli L, Giubilei MA et al (2010) An antigen microarray immunoassay for multiplex screening of mouse monoclonal antibodies. Nat Protoc 5:1932–1944 Dotan N, Altstock RT, Schwarz M et al (2006) Anti-glycan antibodies as biomarkers for diagnosis and prognosis. Lupus 15:442–450 Huflejt ME, Vuskovic M, Vasiliu D et al (2009) Anti-carbohydrate antibodies of normal sera: Findings, surprises and challenges.
Carbohydrate-Based Vaccines: Methods and Protocols by Bernd Lepenies