Monoclonal antibodies to angiotensin converting enzyme from human lung

Danilov, Sergei M.; Allikmets, E. Yu.; Sakharov, Ivan Yu.; Dukhanina, E. A.; Trakht, Ilya

doi:10.1007/bf00842340

Cited by 4 publications

(6 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The boost was carried out by intravenous (IV) injection of antigen without CFA for 3 days. Immediately after, fusion of the spleen cells with myeloma cell line X63.Ag8.653, subclone P3O1, was carried out by a modification of the procedure (24) originally described by Kohler and Milstein (34). Hybridoma supernatants were assayed for anti‐ACE antibodies by enzyme‐linked immunosorbent assay (ELISA) and a plate‐precipitation assay.…”

Section: Methodsmentioning

confidence: 99%

“…A panel of 12 monoclonal antibodies (mAbs) to human lung ACE were developed and characterized (24,25). The panel recognizes at least eight different epitopes on the catalytically active ACE, and has been used to study the structure and function of ACE to deliver genes (26,27) and enzymes to the pulmonary circulation (28,29), and as a diagnostic tool for lung vessel visualization (30).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Monoclonal antibodies to denatured human ACE (CD 143), broad species specificity, reactivity on paraffin sections, and detection of subtle conformational changes in the C‐terminal domain of ACE

et al. 2003

View full text Add to dashboard Cite

Two new mouse monoclonal antibodies (mAbs) were generated to denatured human angiotensin-converting enzyme (ACE, CD143). The clones 2E2 and 3C5, each of the IgG1 kappa chain isotype, detect ACE with high sensitivity, respectively, at 20 ng and 2 ng of protein per lane in Western blotting. They both recognize different epitopes on the C-domain of ACE located between amino acid residues 740 and 992. In formalin-fixed and paraffin-embedded human tissues, immunohistochemistry revealed all known expression sites of ACE, e.g. the epithelial brush borders of proximal kidney tubules, epithelial cells of epididymis, endothelial cells, activated macrophages as well as germ cells during spermatogenesis. In contrast to other mAbs to denatured human ACE, mAbs 2E2 and 3C5 demonstrate cross-reactivity with a broad spectrum of animal species such as monkey, rat, rabbit, cattle, dog, cat, and guinea pig. In addition, mAb 2E2 recognized mouse ACE in Western blotting and on paraffin sections. Our findings suggest that mAbs 2E2 and 3C5 are useful for identifying even subtle changes in ACE conformation resulting from denaturation. These mAbs are also sensitive tools for the detection of minimal amounts of ACE in biological fluids and tissues using proteomics approaches. Their reactivity in routinely processed tissues of various species may prove useful for correlation of ACE expression in animal models to human diseases.

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Monoclonal antibodies to denatured human ACE (CD 143), broad species specificity, reactivity on paraffin sections, and detection of subtle conformational changes in the C‐terminal domain of ACE

et al. 2003

View full text Add to dashboard Cite

show abstract

“…Therefore, during primary screening of hybridomas (generated and cultivated by Elena Allikmets (Sadovnikova) and Ilya Trakht) using ELISA on pure lung ACE we selected ALL positive populations (127), which I tested further on antigen-binding properties in immunocapture test [32], that eliminated possible false-positive (in ELISA) hybridomas with heterofilic antibodies) [33].…”

Section: Generation Of Monoclonal Antibodies To Human Acementioning

confidence: 99%

“…During these 30+ years my lab generated and characterized more than 40 mAbs directed to sequential epitopes (recognizing linear parts of the polypeptide chain) as well as conformational (discontinuous) epitopes on ACE of human, rat, mice and other species [30,32,[34][35][36][51][52][53]. These mAbs were successfully used for ACE quantification using ELISA [18], flow cytometry [11,50,54], Western blotting [34,38], and inmmunohistochemistry [55].…”

Section: Conformational Fingerprinting Of Acementioning

confidence: 99%

“…The initial goal of the project was to obtain mAbs against the organ-specific endothelium, which can be used for targeted drug delivery to different organs. One of the mAbs against ACE, obtained in the first series (9B9), recognized not only human ACE, but also rat ACE [32] (and also hamster and cat ACE, although to a much lesser extent [30], which allowed to trace this mAb after intravenous administration. We found (together with Vladimir Muzykantov) that radioactively labeled mAb 9B9 is extremely effective (up to 50% of the administered dose [73] and specificity (30-50 times more than in other organs and blood) accumulate in the lung [74][75][76].…”

Section: Targeted Drug/gene Delivery Into the Lung Endothelium Using mentioning

confidence: 99%

See 1 more Smart Citation

Conformational Fingerprinting Using Monoclonal Antibodies (on the Example of Angiotensin I-Converting Enzyme-ACE)

Danilov

2017

Mol Biol

Self Cite

View full text Add to dashboard Cite

⎯During the past 30 years my laboratory has generated 40+ monoclonal antibodies (mAbs) directed to structural and conformational epitopes on human ACE as well as ACE from rats, mice and other species. These mAbs were successfully used for detection and quantification of ACE by ELISA, Western blotting, flow cytometry and immunohistochemistry. In all these applications mainly single mAbs were used. We hypothesized that we can obtain a completely new kind of information about ACE structure and function if we use the whole set of mAbs directed to different epitopes on the ACE molecule. When we finished epitope mapping of all mAbs to ACE (and especially, those recognizing conformational epitopes), we realized that we had obtained a new tool to study ACE. First, we demonstrated that binding of some mAbs is very sensitive to local conformational changes on the ACE surface-due to local denaturation, inactivation, ACE inhibitor or mAbs binding or due to diseases. Second, we were able to detect, localize and characterize several human ACE mutations. And, finally, we established a new concept-conformational fingerprinting of ACE using mAbs that in turn allowed us to obtain evidence for tissue specificity of ACE, which has promising scientific and diagnostic perspectives. The initial goal for the generation of mAbs to ACE 30 years ago was obtaining mAbs to organ-specific endothelial cells, which could be used for organ-specific drug delivery. Our systematic work on characterization of mAbs to numerous epitopes on ACE during these years has lead not only to the generation of the most effective mAbs for specific drug/gene delivery into the lung capillaries, but also to the establishment of the concept of conformational fingerprinting of ACE, which in turn gives a theoretical base for the generation of mAbs, specific for ACE from different organs. We believe that this concept could be applicable for any glycoprotein against which there is a set of mAbs to different epitopes.

show abstract

Immunohistochemical study of angiotensin-converting enzyme in human tissues using monoclonal antibodies

et al. 1987

View full text Add to dashboard Cite

The localization of angiotensin-converting enzyme (ACE) in human tissues has been studied by the PAP-method with the use of monoclonal antibody 9 B9 against human lung ACE. The enzyme was detected on the surface of endothelial cells in lung, myocardium, liver, intestine and testis as well as in the epithelial cells of the kidney proximal tubules and intestine. The monoclonal antibody 9 B9 did not react with ACE in the epithelial cells of the testis seminiferous tubules. These data suggest that the antibody 9 B9 recognizes epitope which is shared by the ACE molecule of endothelial cells and renal and intestinal epithelial cells but is not present in testicular ACE, or is not accessible there to the antibody.

show abstract

Monoclonal antibodies to angiotensin converting enzyme from human lung

Cited by 4 publications

References 9 publications

Monoclonal antibodies to denatured human ACE (CD 143), broad species specificity, reactivity on paraffin sections, and detection of subtle conformational changes in the C‐terminal domain of ACE

Monoclonal antibodies to denatured human ACE (CD 143), broad species specificity, reactivity on paraffin sections, and detection of subtle conformational changes in the C‐terminal domain of ACE

Conformational Fingerprinting Using Monoclonal Antibodies (on the Example of Angiotensin I-Converting Enzyme-ACE)

Immunohistochemical study of angiotensin-converting enzyme in human tissues using monoclonal antibodies

Contact Info

Product

Resources

About