PMID- 11170510
OWN - NLM
STAT- MEDLINE
DCOM- 20010426
LR  - 20190921
IS  - 0893-228X (Print)
IS  - 0893-228X (Linking)
VI  - 14
IP  - 1
DP  - 2001 Jan
TI  - Antibody recognition of melphalan adducts characterized using immobilized DNA: 
      enhanced alkylation of G-Rich regions in cells compared to in vitro.
PG  - 71-81
AB  - The bifunctional alkylating agent, melphalan, forms adducts on DNA that are 
      recognized by two previously described monoclonal antibodies, MP5/73 and Amp4/42. 
      Immunoreactivity to MP5/73 was lost when alkylated DNA was exposed to alkaline 
      pH, while Amp4/42 only recognized the structures formed after the alkali 
      treatment. Competitive enzyme-linked immunoadsorbent assays (ELISAs) indicated 
      that in 0.01 and 0.1 M NaOH, loss of immunoreactivity to MP5/73 occurred with 
      half-lives that were at least 2-fold longer than half-lives for gain of 
      immunoreactivity to Amp4/42. This supported previously published evidence that 
      Amp4/42 did not simply recognize all the products formed by alkali treatment of 
      adducts that were initially recognized by MP5/73. Adducts recognized by MP5/73 on 
      RNA were considerably more stable at 100 degrees C and pH 7 than adducts on DNA. 
      This was consistent with the hypothesis that immunorecognition involved N7 
      guanine adducts and ruled out the involvement of phosphotriesters in 
      immunoreactivity. Synthetic oligodeoxyribonucleotides, covalently immobilized 
      onto 96-well plates, were reacted with melphalan and incubated for various 
      periods with alkali, and then the levels of adducts recognized by each antibody 
      in replicate wells were assayed by a direct binding ELISA. Adducts formed on 
      oligodeoxyguanylic acid were recognized very weakly by Amp4/42, unlike other DNA 
      sequences that were tested. Retention of immobilized DNA during alkali treatment 
      was confirmed by immunoassay of cisplatin adducts. Poor recognition by Amp4/42 of 
      adducts in oligodeoxyguanylic acid was confirmed by a competitive ELISA. Amp4/42, 
      unlike MP5/73, efficiently recognized adducts resulting from alkylation of DNA 
      with chlorambucil. It is concluded that the two antibodies recognized melphalan 
      adducts in different DNA sequence environments and that this explains (a) the 
      different alkali stability of immunoreactive adducts and (b) previous results 
      which showed that, in DNA from melphalan-treated cells, adducts recognized by 
      Amp4/42 formed a smaller proportion of total adducts compared to DNA alkylated in 
      vitro. The results presented here indicate that this was caused by a marked 
      cellular influence on the overall sequence-dependent pattern of DNA alkylation or 
      repair.
FAU - McCartney, H
AU  - McCartney H
AD  - Cancer Research Unit and Department of Haematology, Medical School, University of 
      Newcastle upon Tyne, Newcastle upon Tyne NE2 4HH, UK.
FAU - Martin, A M
AU  - Martin AM
FAU - Middleton, P G
AU  - Middleton PG
FAU - Tilby, M J
AU  - Tilby MJ
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Chem Res Toxicol
JT  - Chemical research in toxicology
JID - 8807448
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antineoplastic Agents, Alkylating)
RN  - 0 (Cross-Linking Reagents)
RN  - 0 (DNA Adducts)
RN  - 18D0SL7309 (Chlorambucil)
RN  - 9007-49-2 (DNA)
RN  - Q41OR9510P (Melphalan)
SB  - IM
MH  - Alkylation
MH  - Animals
MH  - Antibodies, Monoclonal/*immunology
MH  - Antineoplastic Agents, Alkylating/*immunology/metabolism
MH  - Base Sequence
MH  - Cattle
MH  - Chlorambucil/chemistry/immunology/metabolism
MH  - Cross-Linking Reagents/chemistry/metabolism
MH  - DNA/chemistry/*immunology/metabolism
MH  - DNA Adducts/chemistry/*immunology
MH  - Drug Stability
MH  - Enzyme-Linked Immunosorbent Assay/methods
MH  - Hot Temperature
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Melphalan/chemistry/*immunology/metabolism
MH  - Molecular Sequence Data
EDAT- 2001/02/15 11:00
MHDA- 2001/05/01 10:01
CRDT- 2001/02/15 11:00
PHST- 2001/02/15 11:00 [pubmed]
PHST- 2001/05/01 10:01 [medline]
PHST- 2001/02/15 11:00 [entrez]
AID - tx000178z [pii]
AID - 10.1021/tx000178z [doi]
PST - ppublish
SO  - Chem Res Toxicol. 2001 Jan;14(1):71-81. doi: 10.1021/tx000178z.