Ephrin type-B receptor 3 (EphB3) binds to transmembrane ephrin-B ligands to regulate cell migration, adhesion, and proliferation. EphB3 exhibits a gradient expression pattern in the normal intestine, with the highest levels at the crypt base, and plays a crucial role in the maintenance of normal intestinal epithelium. Therefore, anti-EphB3 monoclonal antibodies (mAbs) are required for basic research and diagnosis. In this study, we developed novel antihuman EphB3, Eb3Mab-5 (IgG1, κ) and Eb3Mab-11 (IgG1, κ), using the Cell-Based Immunization and Screening (CBIS) method. Eb3Mab-5 and Eb3Mab-11 reacted with EphB3-overexpressed Chinese hamster ovary-K1 (CHO/EphB3) and endogenous EphB3-positive colorectal cancer LS174T in flow cytometry. The apparent binding affinity of Eb3Mab-5 for CHO/EphB3 and LS174T was 7.6 × 10-9 M and 1.7 × 10-8 M, respectively. Eb3Mab-11 could detect EphB3 in western blot analysis and immunohistochemistry. Eb3Mab-5 and Eb3Mab-11, established by the CBIS method, may contribute to the diagnosis and therapy of EphB3-positive tumors.

Androgen receptor (AR) is activated by binding to androgens, which leads to nuclear translocation, dimerization, and binding to androgen response elements (AREs) to regulate gene transcription. AR is important in masculinization during mammalian development and is a major driver of tumor growth in prostate cancer, for which AR pathway inhibitors are the standard treatment. However, the mechanisms by which AR participates in these processes remain unclear. In this study, we describe rat monoclonal antibodies (mAbs) that were generated against human and mouse AR. These mAbs recognize endogenous AR and were shown to be effective in the immunofluorescence staining of human cell lines and mouse tissue sections and in immunoprecipitation experiments. We expect these mAbs to be useful for functional analyses of AR.

CD155, also known as poliovirus receptor (PVR) or Necl-5, is an immunoreceptor with three immunoglobulin-like domains in the extracellular portion and is ubiquitously expressed on the cell surface of hematopoietic and nonhematopoietic cells. Human CD155 contains membrane-bound CD155 (mCD155) and soluble CD155 (sCD155) lacking the transmembrane region encoded by splicing variants of genes. The serum levels of sCD155 increased in patients with a variety of cancers. Furthermore, sCD155 suppresses tumor immunity through the blockade of DNAM-1 signaling. Therefore, sCD155 is potentially helpful for the diagnosis of cancer development and a novel therapeutic target in cancer treatment. However, monoclonal antibodies (mAbs) specific to sCD155, but not mCD155, have not yet been developed. Here, we generated 14 mAbs (named TX119 to TX122 and TX126 to TX135) that recognize the cytoplasmic region of CD155. These mAbs bind to sCD155, but not mCD155, when applied from the outside of the cell. Moreover, we established the assay system to quantify the sCD155 concentration in human serum. Thus, these mAbs can be utilized for the targeting and quantification of sCD155 in the human peripheral blood.

Biological fluids collected from crime scenes play a crucial role in solving serious crimes such as murder, rape, burglary, and theft. Identifying human blood using various methods is crucial for linking disparate pieces of evidence and solving crimes. In this study, the Anti-Human Hemoglobin antibody (Ah-HB) from Sigma-Aldrich® USA was internally validated for human blood identification using the Ouchterlony Double Immunodiffusion (ODD) technique at the DNA and Serology Department of the Punjab Forensic Science Agency in Lahore, Pakistan. Additionally, a comparative analysis was conducted with Seratec® HemDirect (S_HD) strips to evaluate the economic feasibility of both methods. The internal validation included assessments of sensitivity, specificity, and an analysis of real-case work samples to determine the viability of the antibody as a confirmatory test for human blood. Although Ah-HB had lower sensitivity in detecting human blood at higher dilutions (1:10,000), it offered a more cost-effective solution per sample compared with S_HD, which demonstrated higher sensitivity (1:2,000,000) but at a significantly higher cost per sample. Specificity tests revealed no cross-reactivity with nonhuman blood for both Ah-HB and S_HD. This study emphasizes the importance of selecting suitable antibodies for forensic analysis by evaluating sensitivity, specificity, and cost-effectiveness. Ah-HB emerges as a valuable tool for forensic laboratories, providing reliable results at a lower cost compared with S_HD, thereby enhancing the efficiency and effectiveness of criminal investigations.

Erythropoietin-producing hepatocellular receptor A1 (EphA1) is one of the Eph receptor family members, the largest group of receptor tyrosine kinases. EphA1 is expressed in various tissues and regulates cellular homeostasis by interacting with its membrane-bound ephrin ligands and other receptors. EphA1 critically correlates with the pathogenesis in several disorders, including Alzheimer's disease and cancers. Therefore, establishing sensitive monoclonal antibodies (mAbs) for EphA1 has been desired for basic research, diagnosis, and treatment. In this study, a novel specific and sensitive anti-human EphA1 mAb, clone Ea1Mab-30 (mouse IgG1, kappa), was established by the Cell-Based Immunization and Screening (CBIS) method. Ea1Mab-30 demonstrated reactivity with an EphA1-overexpressed Chinese hamster ovary-K1 cell line (CHO/EphA1), an endogenously EphA1-expressing bladder carcinoma cell line (5637), and a colorectal adenocarcinoma cell line (Caco-2) in flow cytometry. Crossreactivities of Ea1Mab-30 with other Eph receptors were not observed. Furthermore, the values of apparent binding affinity for CHO/EphA1 and 5637 were determined to be 8.9 × 10-9 M and 1.7 × 10-9 M, respectively. Furthermore, Ea1Mab-30 detected EphA1 protein in CHO/EphA1 and 5637 lysates using Western blot analysis. Ea1Mab-30 also clearly stained EphA1 of formalin-fixed paraffin-embedded CHO/EphA1 using immunohistochemistry. Ea1Mab-30, established by CBIS method, could help analyze the EphA1-contributed cellular functions and have potential applications in pathological diagnosis and treatment with specificity and high affinity for EphA1-expressing cells.

Immunotherapy, especially monoclonal antibodies, has shown efficacy in modulating the inflammatory response and controlling lipidic pathways, offering new approaches for treating atherosclerotic disease. This article reviews the scientific evidence on the use of therapeutic monoclonal antibodies in treating atherosclerotic disease. An integrative review was carried out using the protocol Whittemore and Knafl framework and the PRISMA 2020 guidelines for systematic reviews. We analyzed clinical trials using monoclonal antibody immunotherapy to treat atherosclerosis. This review used the relevant articles published in Scopus, Embase, PubMed, Cochrane, Web of Science, Scielo, BVS, and Cinhal databases. The period of publication studies that was selected was between 2015 and 2025. Results: 277 articles were identified. One hundred and 22 studies were in duplicate and were excluded. After the complete reading of the studies, only 20 clinical studies were included in this review, both randomized and nonrandomized. Monoclonal antibody immunotherapy presents an innovative approach to treating atherosclerosis by targeting inflammation and lipidic pathway factors. However, these treatments need to be thought out individually, and further research is required to optimize them, minimize risks, and address cost challenges. Combining immunotherapy with other therapies could promote a significant advancement in atherosclerosis management.

One of the most potent therapeutic and diagnostic agents in contemporary medicine is the monoclonal antibody (mAb). mAbs can perform a variety of tasks in breast cancer (BC), including identifying and delivering therapeutic medications to targets, preventing cell development, and suppressing immune system inhibitors including directly attacking cancer cells. mAbs are one of the most effective therapeutic options, particularly for HER2, but they have not been well studied for their use in treating other forms of BC, particularly triple negative breast tumors. Bispecific and trispecific mAbs have created new opportunities for more targeted specific efficacy, which has a positive impact on the viability of antigen specificity. They are more versatile and effective than other forms of treatment, emerging as most popular option for treating BC. However, mAbs have a limit in treatment due to certain adverse effects, including fever, shaking, exhaustion, headache, nausea, and vomiting, as well as rashes, bleeding, and difficulty breathing. To examine the current and prospective future capacities of mAbs with regard to the detection and treatment of BC, the present review highlights advantages and disadvantages of mAb approach.