Hybridoma Technology

Since the development of antibody-production techniques, a number of immunoglobulins have been developed on a large scale using conventional methods. Hybridoma technology opened a new horizon in the production of antibodies against target antigens of infectious pathogens, malignant diseases including autoimmune disorders, and numerous potent toxins. However, these clinical humanized or chimeric murine antibodies have several limitations and complexities. Therefore, to overcome these difficulties, recent advances in genetic engineering techniques and phage display technique have allowed the production of highly specific recombinant antibodies. These engineered antibodies have been constructed in the hunt for novel therapeutic drugs equipped with enhanced immunoprotective abilities, such as engaging immune effector functions, effective development of fusion proteins, efficient tumor and tissue penetration, and high-affinity antibodies directed against conserved targets. Advanced antibody engineering techniques have extensive applications in the fields of immunology, biotechnology, diagnostics, and therapeutic medicines. However, there is limited knowledge regarding dynamic antibody development approaches. Therefore, this review extends beyond our understanding of conventional polyclonal and monoclonal antibodies. Furthermore, recent advances in antibody engineering techniques together with antibody fragments, display technologies, immunomodulation, and broad applications of antibodies are discussed to enhance innovative antibody production in pursuit of a healthier future for humans.

The primary focus of this study was to determine if student
learning outcomes in an undergraduate Statistics course differed
based upon the type of textbook used (e-textbook or
hardcopy). Fifty-six students enrolled in the course were allowed
to choose textbook type. After controlling for student
demographics and academic preparedness, student learning
outcomes for those who used the hardcopy textbook were
significantly higher than for those who used the web-based etextbook.
Though e-textbooks offer many potential benefits
to students, this finding suggests that adoption of e-textbooks
within courses should be guided by the impact on student
learning.

Domoic acid is a potent neurotoxin, glutamate agonist and natural marine biotoxin. It is produced by several diatom algal species of genetically diverse genus Pseudo-nitzschia, red alga Chondria armata and other related species. Historically, widespread domoic acid intoxication incidents have occurred globally. Particularly, in 1987, domoic acid caused an outbreak of human poisoning in Canada by the consumption of contaminated blue mussels (Mytilus edulis) that gave rise to countless clinical illnesses and human deaths. The toxin targets glutamate receptors present in various vital organs and a cascade of ailments characterized the poisoning. The specifically listed neural disorders include memory impairment, coma, recurrent seizures, and epilepsy that led to the name amnesic shellfish poisoning. The occurrence of domoic acid in test samples has been determined by various novel techniques such as liquid chromatography-mass spectrometry, biosensors, enzyme-linked immunosorbent assay and other related hi-tech detection methods. Since the Canada incident, the toxin has intoxicated several marine mammals, sea birds, and contaminated colonized coastal waters around the globe. Subsequently, domoic acid producing algal blooms are accelerating frequently worldwide. Therefore, it poses a global health, safety threat and exposure risks to a significant number of marine and human lives. Genetics of domoic acid producing algal species, molecular markers and therapeutics are currently least known. Therefore, the present review addresses intriguing attributes of domoic acid, such as physicochemical, pharmacological and biological features, sources, and genetic diversity of the producers. In addition, exposure risks including toxicologic pathology of domoic acid poisoning, neurotoxic pathways, detection methods and therapeutics have been inclusively discussed.

Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas which secrete antibodies in the preferred format, species and isotype. Using this platform, we obtained recombinant hybridomas secreting Fab fragments, isotype switched chimeric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemo-enzymatic modification. We believe this versatile platform facilitat...

Nowadays modern information technology’s development is over every field. We often face to a word “hybrid” in our life. Especially, hybrid technologies, hybrid people, hybrid world and etc. At present, it will be better if we enter the concept of hybrid learning system to our scientific world. In XXI century, the word “hybrid” has spread widely. In fact, if the word “hybrid” translated from Latin to English in meaning “seeded”. It has, also, another special meaning “fused”. According to speciality this word is understood diffirently.

Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas, which secrete antibodies in the preferred format, species, and isotype. Using this platform, we obtained recombinant hybridomas secreting Fab′ fragments, isotype-switched chi-meric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemoenzymatic modification. We believe that this versatile platform facilitates antibody engineering for the entire scientific community, empowering preclinical antibody research.

Domoic acid (DA) is a potent toxin, marine biotoxin, and primarily produced by Pseudo-nitzschia. The DA hapten was coupled with bovine serum albumin (BSA), and ovalbumin (OVA) as carrier proteins. DA-BSA conjugate was used as immunogen and DA-OVA as coating antigen. Cell fusion between spleen cells and sp2/0 myeloma cells developed 1C3 hybridoma clone producing 1C3 monoclonal antibody (mAb). Hybridoma was injected into the mice to produce ascites, and further purified by caprylic acid/ammonium sulfate method. The mAb was of IgG3 subclass, and was specific to DA with high affinity (2.5 × 10 8 L/mol). Moreover, western blot exhibited significant specificity to the DA antigens. Indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) showed DA working range of 0.006–0.2 ng/mL. The IC 50 was 0.03 ng/mL with low limit of detection (LOD) of 0.006 ng/mL. Average DA recovery from spiked shellfish extract was 100.56 ± 2.8% with the coefficient variation of 0.01–0.1%. Hence, mAb producing 1C3 hybridoma was successfully developed and could be used to detect DA in contaminated samples.

A monoclonal antibody (mAb) specific to domoic acid which is a potent source of amnesic shellfish poisoning was produced from hybridoma cell line 3G9G1. It was generated by the fusion of Sp2/0 myeloma cells with spleen cells isolated from Balb/c mice with highest-titer 2.50. The mice were immunized with domoic acid-bovine serum albumin. Hybridoma cells were grown on cell culture media supplemented with carbohydrates (CHO) respectively. Monoclonal antibody produced by hybridoma in vitro belongs to the immunoglobulin subtype IgG3 with specific antibody titer 0.65. Culture medium supplemented with maltose was proved to be the most effective for hybridoma clones and achieved best titer at 1.485 as determined by icELISA. The use of supplemented cell culture media is an effective approach for enhanced monospecific-antibody production, better hybridoma growth and viability.

Domoic acid is a highly potent marine toxin which is produced by Pseudo-nitzschia. It causes amnesic shellfish poisoning in humans after consumption of contaminated shellfish. In the present study, hybridoma technology has been used to develop a murine hybridoma cells producing monoclonal antibody by the conditioning of cell culture media with different carbohydrates in vitro. Carbohydrates are important organic enhancements in cell culture media that considerably increase hybridoma cell growth and antibody production.Indirect enzyme-linked immunosorbent assay was established to determine the antibody titers of cell culture supernatants. A hybridoma C4H5 cell line was developed by the fusion of Sp2/0 myeloma cells with spleen cells that were harvested from Balb/c mice immunized with complete DA antigen.The influence of carbohydrates on cell growth and higher protein production of C4H5 cells has been investigated. Galactose, fructose and maltose significantly increased protein production by the factor of 1.7, 2.0 and 2.2, respectively.The study extends understanding of augmented use of superior carbohydrate in cell culture media to improve growth of hybridoma cells and subsequently higher yields of antibody.