[Antibodies to synthetic peptides for the detection of survivin in tumor tissues].
Survivin, an endogenous protein, is a promising marker for the diagnosis of cancer. The aim of the present work was to obtain antibodies specific to survivin and capable of detecting this protein in tumor tissues. Four peptides corresponding to fragments (1-22), (54-74), (80-88)-(153-165), and (118-144) of the survivin-2B sequence were selected and synthesized. Rabbits were immunized with the synthetic peptides. It has been shown that all peptides in a free state, without conjugation with a high-molecular-weight carrier, stimulate the production of antibodies capable of binding with recombinant survivin. Antipeptide antibodies were isolated from sera and their performance in the immunohistochemical detection of survivin in human tumor tissues was studied. It was shown that only antibodies to the (80-88)-(153-165) peptide bind to the survivin present in breast and bladder tumors. The ability of antibodies to this peptide to detect survivin in tumor tissue lysates was demonstrated by immunoblotting. The part of the sequence targeted by the antibodies against the (80-88)-(153-165) peptide was localized using truncated peptide fragments.
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997 Russia. akhidova@gmail.com
E V
EV
Akhidova
T D
TD
Volkova
D O
DO
Koroev
Ia S
IaS
Kim
M P
MP
Filatova
N M
NM
Vladimirova
T A
TA
Karmakova
L E
LE
Zavalishina
Iu Iu
IuIu
Andreeva
O M
OM
Vol'pina
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D014408:Biomarkers, Tumor / Q000032:analysis; D001943:Breast Neoplasms / Q000737:chemistry*; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000276:immunology*; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D000077022:Survivin; D001749:Urinary Bladder Neoplasms / Q000737:chemistry*
D000906:Antibodies; C107489:BIRC5 protein, human; D014408:Biomarkers, Tumor; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009842:Oligopeptides; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000077022:Survivin