Detection of complement activation using monoclonal antibodies against C3d

Thurman, Joshua M.; Kulik, Liudmila; Orth, Heather; Wong, M. A.; Renner, Brandon; Sargsyan, S. Anna; Mitchell, Lynne M.; Hourcade, Dennis E.; Hannan, Jonathan P.; Kovacs, Julie A.; Coughlin, Beth; Woodell, Alex; Pickering, Matthew C.; Rohrer, Bärbel; Holers, V. Michael

doi:10.1172/jci65861

Cited by 79 publications

(103 citation statements)

References 45 publications

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“…Selected druglike compounds were obtained from Enamine, Pharmeks, ChemDiv, ChemBridge, and Maybridge. Mols 4,5,16,36,42, and 68 (ChemBridge) were assessed for purity by LC−MS. The specific purities for each molecule are 91.4% (Mol4), 91.4% (Mol5), 98.4% (Mol16), 97.7% (Mol36), 95.0% (Mol42), and 88.7% (Mol68).…”

Section: ■ Discussionmentioning

confidence: 99%

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Discovery of Small Molecules for Fluorescent Detection of Complement Activation Product C3d

et al. 2015

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Complement activation plays a major role in many acute and chronic inflammatory conditions. C3d, a terminal product of complement activation, remains covalently attached to cells and is an excellent biomarker of complementmediated inflammation. We employed a virtual highthroughput screening protocol to identify molecules with predicted binding to complement C3d and with intrinsic fluorescence properties to enable detection. Pharmacophore models were developed based on known C3d−ligand interactions and information from computational analysis of structural and molecular dynamics data. Iterative pharmacophore-based virtual screening was performed to identify druglike molecules with physicochemical similarity to the natural C3d ligand CR2. Hits from the pharmacophore screens were docked to C3d and ranked based on predicted binding free energies. Top-ranked molecules were selected for experimental validation of binding affinity to C3d, using microscale thermophoresis, and for their suitability to become molecular imaging agents, using fluorescence spectroscopy. This work serves as a foundation for identifying additional fluorescent molecules with high-affinity for C3d that will subsequently be explored as noninvasive in vivo diagnostics of complementmediated inflammation, for spatiotemporal monitoring of disease progression, and for targeting therapeutics to sites of inflammation.

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Section: ■ Discussionmentioning

confidence: 99%

“…2 However, measurement of C3d levels typically requires tissue biopsies. 5,6,9,15 It is therefore desirable to develop a noninvasive method to detect levels of complement activation fragments on tissues.…”

Section: ■ Introductionmentioning

confidence: 99%

Discovery of Small Molecules for Fluorescent Detection of Complement Activation Product C3d

et al. 2015

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“…Because CR2 does not bind intact plasma C3 it can be used to target therapeutic and diagnostic agents to sites of complement activation 8, 78– 80 . We have also developed several monoclonal antibodies to C3d that do not bind to intact C3 or to C3b 81 . These antibodies bind C3d with a high affinity (<1 nM) and target sites of complement activation when injected systemically 81 .…”

Section: The Promise Of Molecular Imaging For Monitoring Ln and Npslementioning

confidence: 99%

“…We have also developed several monoclonal antibodies to C3d that do not bind to intact C3 or to C3b 81 . These antibodies bind C3d with a high affinity (<1 nM) and target sites of complement activation when injected systemically 81 .…”

Section: The Promise Of Molecular Imaging For Monitoring Ln and Npslementioning

confidence: 99%

“…As outlined above, PET probes can be detected with higher sensitivity than SPIONs ( Figure 1), and we have developed high-affinity anti-C3d monoclonal antibodies that accumulate at sites of complement activation after systemic injection 81 . Factor H deficient ( fH -/- ) mice develop spontaneous glomerulonephritis characterized by abundant glomerular C3 fragments 83 .…”

Section: The Promise Of Molecular Imaging For Monitoring Ln and Npslementioning

confidence: 99%

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Non-invasive imaging to monitor lupus nephritis and neuropsychiatric systemic lupus erythematosus

Thurman

Serkova

2015

F1000Res

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Systemic lupus erythematosus (SLE) is an autoimmune disease that can affect multiple different organs, including the kidneys and central nervous system (CNS). Conventional radiological examinations in SLE patients include volumetric/ anatomical computed tomography (CT), magnetic resonance imaging (MRI) and ultrasound (US). The utility of these modalities is limited, however, due to the complexity of the disease. Furthermore, CT and MRI contrast agents are contraindicated in patients with renal impairment. Various radiologic methods are currently being developed to improve disease characterization in patients with SLE beyond simple anatomical endpoints. Physiological non-contrast MRI protocols have been developed to assess tissue oxygenation, glomerular filtration, renal perfusion, interstitial diffusion, and inflammation-driven fibrosis in lupus nephritis (LN) patients. For neurological symptoms, vessel size imaging (VSI, an MRI approach utilizing T2-relaxing iron oxide nanoparticles) has shown promise as a diagnostic tool. Molecular imaging probes (mostly for MRI and nuclear medicine imaging) have also been developed for diagnosing SLE with high sensitivity, and for monitoring disease activity. This paper reviews the challenges in evaluating disease activity in patients with LN and neuropsychiatric systemic lupus erythematosus (NPSLE). We describe novel MRI and positron-emission tomography (PET) molecular imaging protocols using targeted iron oxide nanoparticles and radioactive ligands, respectively, for detection of SLE-associated inflammation.

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Fluorescence Imaging

Albani

2016

Encyclopedia of Analytical Chemistry

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A number of fluorescence imaging techniques show diagnostic promise. Imaging endogenous fluorescence has been proposed as a method for cancer diagnosis. Unfortunately, tissue auto fluorescence is relatively weak and poor contrast between malignant and normal tissue is seen. Contrast may be enhanced with the addition of fluorescent materials that are selectively accumulated by malignant cells, such as fluorescein or porphyrin derivatives. The limited penetration of light at the emission maxima of these materials restricts the use of fluorescence techniques utilizing these chromophores to superficial phenomena. However, many potential applications still exist. For example, monitoring fluorescence during surgery may allow resection margins to be clearly delineated. Other exogenous chromophores that may have diagnostic utility include indocyanine green (ICG). Techniques based on visualization of the distribution of ICG fluorescence (i.e., choroidal angiography) are already prominent in ophthalmology. ICG fluorescence imaging may also find a useful niche in monitoring of burns and transplant tissue. In addition, monitoring of vascular parameters during cardiac surgery presents exciting opportunities. For example, low oxygen levels (e.g., during bypass surgery) in the heart can result in alterations in microvascular permeability. As ICG is largely bound to serum albumin, it should not be seen in extravascular spaces in normal hearts. However, increased permeability will allow albumin to diffuse into the extravascular spaces, and diffuse fluorescence across the surface of the heart will be seen. In principle, the infusion of polymers (e.g., dextrans) of various molecular weights labeled with dyes that fluoresce at various wavelengths will allow the assessment of the porosity of capillary beds in such systems. Immunofluorescence techniques have the potential to provide unmatched sensitivity and specificity. The unique nature of antibody‐antigen interactions ensures specific delivery of the fluorophore to the site of interest. The specific interaction of labeled antibodies with antigens means that the fluorophore persists in the body for a prolonged period of time (days). Following a single injection of labeled antibody, repeated measurements on the same site over the course of hours or days allow kinetic information to be readily obtained. In principle, this means that the effect of therapeutic intervention, i.e., radiation therapy, chemotherapy, and so on, can be monitored.

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Detection of complement activation using monoclonal antibodies against C3d

Cited by 79 publications

References 45 publications

Discovery of Small Molecules for Fluorescent Detection of Complement Activation Product C3d

Discovery of Small Molecules for Fluorescent Detection of Complement Activation Product C3d

Non-invasive imaging to monitor lupus nephritis and neuropsychiatric systemic lupus erythematosus

Fluorescence Imaging

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