Generation, expression and utilization of single-domain antibodies for in vivo protein localization and manipulation in sea urchin embryos

Schrankel, Catherine S.; Gökirmak, Tufan; Lee, Chang-Wook; Chang, Geoffrey; Hamdoun, Amro

doi:10.1016/bs.mcb.2018.11.005

Cited by 7 publications

(5 citation statements)

References 40 publications

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“…Sharks were tagged either on the first dorsal fin with a plastic Rototag (Premier 1) or just below the first dorsal fin with a metal dart tag. The tag type and location depended both on species and size of shark at capture (Kohler and Turner, 2001). Catch data were converted to catch per unit effort (CPUE), expressed as individuals 100 hooks −1 hour −1 .…”

Section: Bottom Longline Surveymentioning

confidence: 99%

Defining Sex-Specific Habitat Suitability for a Northern Gulf of Mexico Shark Assemblage

et al. 2020

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Understanding the factors that influence species' distributions is crucial for implementing effective management and conservation practices, yet difficult for highly vagile species like sharks. Many shark species demonstrate either spatial and/or temporal sexual segregation, further confounding accurate quantification of habitat suitability. Given the importance of understanding spatiotemporal patterns in the distribution of coastal shark assemblages, we sought to quantify sex-specific abiotic factors that influence seasonal variation in a coastal shark assemblage using data from a long-term fisheriesindependent bottom longline program in the northern Gulf of Mexico. Catch data (individuals/100 hooks/hour) were coupled with a suite of potentially predictive variables: surface and bottom values for temperature (• C) and salinity (psu), sea surface height (m), three-dimensional surface and bottom current velocity (u, v, w, in m/s), bottom dissolved oxygen (mg/l), depth (m), substrate grain size (mm), daylength (min), and distance from shore (km). Data were analyzed using boosted regression trees (BRT) to describe the relationships between catch data and environmental factors potentially influencing sex-specific species distribution and abundance. Between May 2006 and November 2018, we conducted 1,226 bottom longline sets and caught 13,742 individuals encompassing 67 species. The majority of the animals captured (74%) were elasmobranchs, primarily sharks. Two species from each of the following three categories were selected for further analyses: small coastal sharks (Atlantic sharpnose shark Rhizoprionodon terraenovae and blacknose shark Carcharhinus acronotus), large coastal sharks (blacktip shark C. limbatus and sandbar shark C. plumbeus), and shelfassociated sharks (smoothhound sharks, Mustelus spp. and scalloped hammerhead Sphyrna lewini). Depth and distance from shore were the strongest predictors of distribution and relative abundance, followed by longitude and bottom salinity; other factors (e.g., temperature, daylength, substrate grain size) were less predictive. For the six species examined, predictive factors were often the same for males and females, although the range of preferred values varied. Surprisingly, the importance of these

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Section: Bottom Longline Surveymentioning

confidence: 99%

Defining Sex-Specific Habitat Suitability for a Northern Gulf of Mexico Shark Assemblage

et al. 2020

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“…Moreover, the recent increase in the use of phage display antibody libraries has proven the advantages of using sdAb fragments in the development of therapeutic antibodies for cancer treatment. Single domain antibodies are characterized for their smaller size while keeping all the binding properties of full-length antibodies [ 49 ]. In this study anti-TK1 sdAbs were isolated from a sdAb library through phage display.…”

Section: Discussionmentioning

confidence: 99%

Selection of human single domain antibodies (sdAb) against thymidine kinase 1 and their incorporation into sdAb-Fc antibody constructs for potential use in cancer therapy

et al. 2022

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Thymidine Kinase 1 (TK1) is primarily known as a cancer biomarker with good prognostic capabilities for both hematological and solid malignancies. However, recent studies targeting TK1 at protein and mRNA levels have shown that TK1 may be useful as a therapeutic target. In order to examine the use of TK1 as a therapeutic target, it is necessary to develop therapeutics specific for it. Single domain antibodies (sdAbs), represent an exciting approach for the development of immunotherapeutics due to their cost-effective production and higher tumor penetration than conventional antibodies. In this study, we isolated sdAb fragments specific to human TK1 from a human sdAb library. A total of 400 sdAbs were screened through 5 rounds of selection by monoclonal phage ELISA. The most sensitive sdAb fragments were selected as candidates for preclinical testing. The sdAb fragments showed specificity for human TK1 in phage ELISA, Western blot analysis and had an estimated limit of detection of 3.9 ng/ml for the antibody fragments 4-H-TK1_A1 and 4-H-TK1_D1. The antibody fragments were successfully expressed and used for detection of membrane associated TK1 (mTK1) through flow cytometry on cancer cells [lung (~95%), colon (~87%), breast (~53%)] and healthy human mononuclear cells (MNC). The most sensitive antibody fragments, 4-H-TK1_A1 and 4-H-TK1_D1 were fused to an engineered IgG1 Fc fragment. When added to cancer cells expressing mTK1 co-cultured with human MNCs, the anti-TK1-sdAb-IgG1_A1 and D1 were able to elicit a significant antibody-dependent cell-mediated cytotoxicity (ADCC) response against lung cancer cells compared to isotype controls (P<0.0267 and P<0.0265, respectively). To our knowledge this is the first time that the isolation and evaluation of human anti-TK1 single domain antibodies using phage display technology has been reported. The antibody fragments isolated here may represent a valuable resource for the detection and the targeting of TK1 on tumor cells.

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“…Moreover, the recent increase in the use of phage display antibody libraries have proven the advantages of using sdAb fragments in the development of therapeutic antibodies for cancer treatment. Single domain antibodies are characterized for their smaller size while keeping all the binding properties of full-length antibodies [47]. In this study anti-TK1 sdAbs were isolated from a sdAb library through phage display.…”

Section: Discussionmentioning

confidence: 99%

Selection of Human Single Domain Antibodies (sdAb) Against Thymidine Kinase 1 and Their Incorporation Into sdAb-Fc Antibody Constructs For Potential Use In Cancer Therapy

et al. 2021

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Thymidine Kinase 1 (TK1) is primarily known as a cancer biomarker with good prognostic capabilities for liquid and solid malignancies. However, recent studies targeting TK1 at protein and mRNA levels have shown that TK1 may be useful as a tumor target. In order to examine the use of TK1 as a tumor target, it is necessary to develop therapeutics specific for TK1. Single domain antibodies (sdAbs), represent an exciting approach for the development of immunotherapeutics due to their cost-effective production and higher tumor penetration than conventional antibodies. In this study, we isolated sdAb fragments specific to human TK1 from a human sdAb library. A total of 400 sdAbs were screened through 5 rounds of selection by monoclonal phage ELISA. The most sensitive sdAb fragments were selected as candidates for preclinical testing. The sdAb fragments showed specificity for human TK1 in phage ELISA, Western blot analysis and had a limit of detection of 3.9 ng/ml for 4-H-TK1_A1 and 1.9 ng/ml for 4-H-TK1_D1. The antibody fragments were successfully expressed and used for detection of membrane associated TK1 (mTK1) through flow cytometry on cancer cells [lung (~95%), colon (~87%), breast (~53%)] and healthy human mono nuclear cells (MNC). The most sensitive antibody fragments, 4-H-TK1_A1 and 4-H-TK1_D1 were fused to an engineered IgG1 Fc fragment. When added to cancer cells expressing mTK1 co-cultured with human MNC, the anti-TK1-sdAb-IgG1_A1 and D1 were able to elicit a significant antibody-dependent cell-mediated cytotoxicity (ADCC) response by human MNCs against lung cancer cells compared to isotype controls (P<0.0267 and P<0.0265, respectively). To our knowledge this is the first time that the isolation and evaluation of human anti TK1 single domain antibodies using phage display technology has been reported. The antibody fragments isolated here may represent a valuable resource for the detection and the targeting of TK1 in tumor cells

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Generation, expression and utilization of single-domain antibodies for in vivo protein localization and manipulation in sea urchin embryos

Cited by 7 publications

References 40 publications

Defining Sex-Specific Habitat Suitability for a Northern Gulf of Mexico Shark Assemblage

Defining Sex-Specific Habitat Suitability for a Northern Gulf of Mexico Shark Assemblage

Selection of human single domain antibodies (sdAb) against thymidine kinase 1 and their incorporation into sdAb-Fc antibody constructs for potential use in cancer therapy

Selection of Human Single Domain Antibodies (sdAb) Against Thymidine Kinase 1 and Their Incorporation Into sdAb-Fc Antibody Constructs For Potential Use In Cancer Therapy

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