Strategy for Automated Development of Reversed‐Phase HPLC Methods for Domain‐Specific Characterization of Monoclonal Antibodies

2021 ◽  
pp. 177-198
Author(s):  
Jennifer La ◽  
Mark Condina ◽  
Leexin Chong ◽  
Craig Kyngdon ◽  
Matthias Zimmermann ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Reversed Phase ◽  
Reversed Phase Hplc ◽  
Domain Specific

Characterization of in vitro proteolytic processing of β-endorphin by reversed-phase HPLC

Peptides ◽  
1984 ◽  
Vol 5 (6) ◽  
pp. 1037-1042 ◽  
Author(s):  
Thomas P. Davis ◽  
Hans Schoemaker ◽  
Alison J. Culling-Berglund
Keyword(s):  
Reversed Phase ◽  
Proteolytic Processing ◽  
Reversed Phase Hplc

scholarly journals Purification and characterization of an L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT)-specific suppressor factor from genetic responder mice.

1983 ◽  
Vol 158 (4) ◽  
pp. 1034-1047 ◽  
Author(s):  
C M Sorensen ◽  
C W Pierce ◽  
D R Webb
Keyword(s):  
Specific Activity ◽  
Reversed Phase ◽  
Biologically Active ◽  
Reversed Phase Hplc ◽  
Purification And Characterization ◽  
Suppressor Factor ◽  
Chemical Homogeneity ◽  
T Cell Factor ◽  
Molecular Weights

A hybridoma-derived, GAT-specific suppressor T cell factor (GAT-TsFR) from responder C57BL/10 mice has been purified to apparent chemical homogeneity using reversed phase HPLC techniques. 40 l of starting material yielded approximately 880 micrograms protein with a specific activity of 28.4 X 10(3) S50 U/ng protein representing a purification factor of 4.2 X 10(6). Purified GAT-TsFR is a hydrophobic protein with a minimum molecular weight of 18,000 that is capable of forming biologically active aggregates with molecular weights of 28,000, 64,000 and approximately 84,000 and has a pI of 6.4. GAT-TsFR is a glycoprotein that binds GAT and GT, but not GA, and bears determinants encoded by the I-J subregion of the H-2 complex. This GAT-TsFR derived from an H-2b responder haplotype to GAT is compared with GAT-TsF derived from the nonresponder H-2q haplotype on the basis of biochemical and some serological properties.

Characterization of Highly Polar Dna Adducts Derived from Dibenz[A,H]Anthracene (DBA), 3,4-Dihydroxy-3,4-Dihydro-DBA, and 3,4,10,11-Tetrahydroxy-3,4,10,11-Tetrahydro-DBA

1993 ◽  
Vol 9 (3) ◽  
pp. 503-509 ◽  
Author(s):  
Juergen Fuchs ◽  
Jiri Mlcoch ◽  
Franz Oesch ◽  
Karl Ludwig Platt
Keyword(s):  
Dna Adducts ◽  
Liver Microsomes ◽  
Metabolic Activation ◽  
Reversed Phase ◽  
Reversed Phase Hplc ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Calf Thymus

Two highly polar DNA adducts were found after metabolic activation of 3,4,10,11-tetrahydroxy-3,4,10,11-tetrahydrodibenz[ a,h]anthracene(DBA-3,4;10, 11-bisdiol) by liver microsomes isolated from male Sprague-Dawley rats pretreated with Aroclor 1254 in presence of calf thymus DNA. These DNA adducts could be assigned to the metabolites of dibenz[ a,h]anthracene (DBA), of 3R,4R,10R,11R-tetrahydroxy-3,4,10,11-tetrahydro-DBA and of 3R,4R,10S,US-tetrahydroxy-3,4,10,11-tetrahydro-DBA. DNA adducts derived from metabolites of 3S,4S,10S,11S-tetrahydroxy-3,4,10,11-tetrahydro-DBA were not found. These highly polar adducts also could be detected by reversed phase HPLC after incubation of dibenz[ a,h]anthracene, 3R,4R-dihydroxy-3,4-dihydro-DBA ((-)-DBA-3,4-diol) and 3S,4S- dihydroxy-3,4-dihydro-DBA ((+)-DBA-3,4-diol) with DNA in presence of the activating system. After incubation of 14C labelled DBA DNA adducts derived from DBA-3,4;10,11-bisdiol were found in a fraction of 38% and bay region 3,4-dihydroxy-1,2-epoxy-1,2,3,4-tetrahydro-DBA-DNA adducts at a level of 25%. DBA-3,4; 10,11-bisdiol exhibited a higher DNA binding yield (38 × 12 pmollmg DNA) than (-)-DBA-3,4-diol (23 × 6 pmol/mg DNA), the most mutagenic 3,4-diol enantiomer. For (+)-DBA-3,4-diol the highly polar DNA adducts derived from DBA-3,4;10,11-bisdiol were by far the most predotmnant adducts in vitro.

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