Human Antibodies - Volume 26, issue 1

Authors: Shanehbandi, Dariush | Majidi, Jafar | Kazemi, Tohid | Baradaran, Behzad | Aghebati-Maleki, Leili

Article Type: Research Article

Abstract: BACKGROUND: CD20-based targeting of B-cells in hematologic malignancies and autoimmune disorders is associated with outstanding clinical outcomes. Isolation and characterization of VH and VL cDNAs encoding the variable regions of the heavy and light chains of monoclonal antibodies (MAb) is necessary to produce next generation MAbs and their derivatives such as bispecific antibodies (bsAb) and single-chain variable fragments (scFv). OBJECTIVE: This study was aimed at cloning and characterization of the VH and VL cDNAs from a hybridoma cell line producing an anti-CD20 MAb. METHODS: VH and VL fragments were amplified, cloned and characterized. …Furthermore, amino acid sequences of VH, VL and corresponding complementarity-determining regions (CDR) were determined and compared with those of four approved MAbs including Rituximab (RTX), Ibritumomab tiuxetan, Ofatumumab and GA101. RESULTS: The cloned VH and VL cDNAs were found to be functional and follow a consensus pattern. Amino acid sequences corresponding to the VH and VL fragments also indicated noticeable homologies to those of RTX and Ibritumomab. Furthermore, amino acid sequences of the relating CDRs had remarkable similarities to their counterparts in RTX and Ibritumomab. CONCLUSIONS: Successful recovery of VH and VL fragments encourages the development of novel CD20 targeting bsAbs, scFvs, antibody conjugates and T-cells armed with chimeric antigen receptors. Show more

Keywords: CD20, targeted therapy, monoclonal antibody, bispecific antibody, chimeric antigen receptor, antibody drug conjugate

DOI: 10.3233/HAB-170314

Citation: Human Antibodies, vol. 26, no. 1, pp. 1-6, 2018

Authors: Amanzadeh, Amir | Heidarnejad, Fatemeh | Abdollahpour-Alitappeh, Meghdad | Molla-Kazemiha, Vahid | Yari, Shamsi | Hadizadeh-Tasbiti, Alireza | Habibi-Anbouhi, Mahdi | Abolhassani, Mohsen | Shokrgozar, Mohammad Ali

Article Type: Research Article

Abstract: AIM: CD44s antigens have been suggested as an efficient biomarker for cancer stem cells. Current study aimed to develop a hybridoma that producing a high affinity murine anti-human CD44 monoclonal antibody for early diagnostic laboratory tests of some cancer. MATERIALS AND METHODS: To make hybridoma against CD44, mice were immunized with MDA-MB-468 cells. Resulted hybridomas using three culture media were screened by indirect ELISA, then cloned by limiting dilution, and isotype was determined after obtaining ascitic fluid and antibody purification. RESULTS: We obtained a stable secreting clone, capable of secreting a high-affinity monoclonal …antibody against CD44 protein, IgG2a kappa, with the affinity of 5.4 × 10-8 M without cross-reactivity. CONCLUSION: We could establish a hybridoma in a native form. This stable and high-affinity anti-CD44 mAb has a potential for diagnostic procedures and laboratory research. Thus, it could be exploited as a suitable tool for target-specific diagnosis and even treatment in several cancers. Show more

Keywords: CD44 monoclonal antibody, biomarker, high-affinity, hybridoma, ascitic fluid

DOI: 10.3233/HAB-170315

Citation: Human Antibodies, vol. 26, no. 1, pp. 7-15, 2018

Authors: Sayad, Arezou | Kelarijani, Mohsen Khakzad | Sajjadi, Elham | Taheri, Mohammad

Article Type: Research Article

Abstract: BACKGROUND: Multiple sclerosis (MS) as an auto-immune disease is an inflammatory, demyelinating disease of the central nervous system. Certain genes have shown to be involved in the initiation of MS but the specific role of some of them, e.g. IFNAR1 has not been identified in certain populations yet. OBJECTIVE: The IFNAR1 as a type I membrane protein shapes one of the two chains of a receptor for interferons alpha and beta. METHODS: To find out how IFNAR1 functions in the Iranian population, the researchers compared the expression level of this …gene in relapsing-remitting MS (RR-MS) samples with normal individuals. RNA from the whole blood of 50 RR-MS patients and 50 normal controls were extracted. All patients were HLA-DRB1*15 negative and were responders to interferon-beta with a normal vitamin D level. The level of IFNAR1 gene expression was measured using quantitative RT-PCR. RESULTS: According to the results the RR-MS patients manifested a statistically higher expression level of IFNAR1 than their normal counterparts (p= 0.012). Age-wise, females between the ages, 30 to 40 had a significant increase (p= 0.046) but males under 30 showed a statistically meaningful decrease in the expression level (p= 0.04). In terms of sex, only the female patients manifested a statistically significant increase in IFNAR1 (p= 0.004). CONCLUSIONS: The overall results show an increase in IFNAR1 level in MS patients treated with IFN-B. Show more

Keywords: IFNAR1, expression, multiple sclerosis, real time PCR

DOI: 10.3233/HAB-170316

Citation: Human Antibodies, vol. 26, no. 1, pp. 17-22, 2018

Authors: Salim, Masome Afiati | Eftekharian, Mohammad Mahdi | Taheri, Mohammad | Yousef Alikhani, Mohammad

Article Type: Research Article

Abstract: BACKGROUND: Multiple sclerosis (MS) is a chronic autoimmune disease that disables central nervous system (CNS) system. Cytomegalovirus (CMV) probably has an important role in the MS pathology. The infection with helicobacter pylori also is recognized as a protective agent against MS in female. METHODS: Serum samples were isolated and frozen at - 70 ∘ C. The earlier mentioned anti-virus antibodies and antibacterial antibodies were quantified by Elisa kit. RESULTS: The results showed that IgG antibody average value against cytomegalovirus in the blood of multiple sclerosis patients not only …decreased but also was significant statistically (p < 0.05). IgM and IgG antibodies average value in the blood of multiple sclerosis patients against helicobacter pylori shown a statistically significant decrease (p < 0.05). CONCLUSION: Therefore it may be considered that probably helicobacter pylori presence in the individuals especially in female can alleviate MS signs. CMV infection can intensify the symptoms in multiple sclerosis patients. Show more

Keywords: Multiple sclerosis, helicobacter pylori, cytomegalovirus, ELISA, Hamadan

DOI: 10.3233/HAB-170317

Citation: Human Antibodies, vol. 26, no. 1, pp. 23-28, 2018

Authors: Kuhn, Philipp | Thiem, Stefanie | Steinert, Michael | Purvis, Duncan | Lugmayr, Veronika | Treutlein, Ulrich | Plobner, Lutz | Leiser, Robert-Matthias | Hust, Michael | Dübel, Stefan

Article Type: Research Article

Abstract: Legionella are Gram-negative bacteria that are ubiquitously present in natural and man-made water reservoirs. When humans inhale aerosolized water contaminated with Legionella , alveolar macrophages can be infected, which may lead to a life-threatening pneumonia called Legionnaires’ disease. Due to the universal distribution of Legionella in water and their potential threat to human health, the Legionella concentration in water for human use must be strictly monitored, which is difficult since the standard detection still relies on lengthy cultivation and analysis of bacterial morphology. In this study, an antibody against L. pneumophila has been generated from the …naïve human HAL antibody libraries by phage-display for the first time. The panning was performed on whole bacterial cells in order to select antibodies that bind specifically to the cell surface of untreated Legionella . The bacterial cell wall component lipopolysaccharide (LPS) was identified as the target structure. Specific binding to the important pathogenic L. pneumophila strains Corby, Philadelphia-1 and Knoxville was observed, while no binding was detected to seven members of the families Enterobacteriaceae , Pseudomonadaceae or Clostridiaceae . Production of this antibody in the recombinant scFv-Fc format using either a murine or a human Fc part allowed the set-up of a sandwich-ELISA for detection of Legionella cells. The scFv-Fc construct proved to be very stable, even when stored for several weeks at elevated temperatures. A sensitivity limit of 4,000 cells was achieved. The scFv-Fc antibody pair was integrated on a biosensor, demonstrating the specific and fast detection of L. pneumophila on a portable device. With this system, 10,000 Legionella cells were detected within 35 min. Combined with a water filtration/concentration system, this antibody may be developed into a promising reagent for rapid on-site Legionella monitoring. Show more

Keywords: Phage display, panning, antibody engineering, antibody fragments, Legionella, LPS, biosensor, diagnostics

DOI: 10.3233/HAB-170318

Citation: Human Antibodies, vol. 26, no. 1, pp. 29-38, 2018

Authors: Voets, Philip J.G.M.

Article Type: Research Article

Abstract: Despite its relevance to many biomedical fields, relatively little effort has been put into a comprehensible quantitative description of the effect of reaction temperature on the interaction between antigens and their antibodies. In this article, a novel, straightforward mathematical model is proposed, which aims to describe the effect of temperature on antigen-antibody kinetics. The model proposed in this article could hopefully provide clinicians, immunologists, and biochemists with an improved insight into the kinetic effect of fluctuations in reaction temperature on antigen-antibody-dependent processes and therefore into the kinetics of the humoral adaptive immune response.

Keywords: Antigen, antibody, kinetics, temperature, mathematical model

DOI: 10.3233/HAB-170319

Citation: Human Antibodies, vol. 26, no. 1, pp. 39-41, 2018