No abstract available.
Humans* ; Hybridomas*

No abstract available.
Cryptococcosis* ; Hybridomas* ; Immunity, Cellular* ; T-Lymphocytes*

No abstract available.
Cryptococcosis* ; Hybridomas* ; Immunity, Cellular* ; T-Lymphocytes*

Mouse hybridoma monoclonal antibody is the most commonly used antibody in immunology because of its stable source, easy preparation in later stage and high yield. The traditional time-consuming and laborious hybridoma preparation technology could not meet the growing market demand. In this paper, we describe the rapid preparation techniques involved in antigen design and screening, B cell enrichment and screening, transgenic myeloma cells, fusion technology improvement, positive hybridoma cell screening and rapid detection of monoclonal antibody performance, to provide a reference for rapid preparation of mouse hybridoma monoclonal antibody.
Animals ; Antibodies, Monoclonal ; Antigens ; B-Lymphocytes ; Hybridomas ; Mice

The 21st century is regarded as the century of biotechnological drugs, among which monoclonal antibodies and their derived targeting drugs have established themselves as the leading modality of biopharmaceutical pharmaceutics for a wide range of indications covering malignant tumors and autoimmune disorders. Since the manufacturing of the first antibody drug from hybridoma cells, the technologies have been intensely studied and there emerged numerous breakthroughs in recombinant cell line establishment, antibody expression and purification, quality control and other related areas. This article summarizes the critical progresses of antibody drugs expression technologies, especially of mammalian cell expression system, Escherichia coli expression system, the transgenic animal reactor and the cell free protein synthesis system, to give a detailed illustration of the recent advances in antibody drugs development.
Animals ; Antibodies, Monoclonal ; Biotechnology ; Cell Line ; Escherichia coli ; Hybridomas

Anti-idiotype antibody (anti-id Ab) which recognizes idiotope in the variable region of immunoglobulin (Ig) can regulate Ab production by B cells in vivo and in vitro. Although it has been reported that anti-id Ab can suppress IgM production by lymphocytes or hybridoma cells without suppression of cell proliferation, the regulatory mechanism of anti-id Ab is not completely understood. We studied the effects of anti-id Ab on the production of IgG class anti-DNA Ab by hybridoma cells, on the proliferation of cells, and on the transcription levels of Ig genes. In contrast to suppressive effect of anti-id Ab on the production of IgM previously reported by others, stimulatory effects of anti-id Ab on the production of IgG by hybridoma cells as well as the proliferation of these .cells were observed. However, little effect of anti-id Ab on the transcription levels of Ig genes was observed. These results suggest that anti-id Ab can increase Ab production by stimulation of cell proliferation. Furthermore, these results suggest that the effect of anti-id Ab on the production of Ab may be determined by the difference in class of Ab produced by hybridoma cells following the treatment with anti-id Ab.
Antibody Formation* ; B-Lymphocytes ; Cell Proliferation ; DNA* ; Genes, Immunoglobulin ; Hybridomas* ; Immunoglobulin G ; Immunoglobulin M ; Immunoglobulins ; Lymphocytes

The follicle-associated epithelium (FAE) of Peyer's patches (PPs) contains M cells that are important for reducing mucosal immune responses by transporting antigens into the underlying lymphoid tissue. We generated a monoclonal antibody (C6) that reacted with the FAE of calf ileal PPs, and analyzed the characteristics of C6 using immunohistochemistry and Western blotting. FAE of the ileal PP was stained with C6 during both late fetal developmental and postnatal stages. Neither the villous epithelial cell nor intestinal crypt basal cells were stained at any developmental stage. During the prenatal stages, FAE of the jejunal PP was C6-negative. However, a few C6-positive cells were distributed diffusely in some FAE of the jejunal PPs during the postnatal stages. The protein molecular weight of the antigen recognized by C6 was approximately 45 kDa. These data show that C6 is useful for identifying the FAE in ileal PPs and further suggest that differentiation of the FAE in these areas is independent of external antigens.
Animals ; Antibodies, Monoclonal/*immunology ; *Cattle ; Fetus ; Hybridomas ; Ileum/*ultrastructure ; Intestinal Mucosa/*immunology ; Peyer's Patches/*immunology/ultrastructure

The follicle-associated epithelium (FAE) of Peyer's patches (PPs) contains M cells that are important for reducing mucosal immune responses by transporting antigens into the underlying lymphoid tissue. We generated a monoclonal antibody (C6) that reacted with the FAE of calf ileal PPs, and analyzed the characteristics of C6 using immunohistochemistry and Western blotting. FAE of the ileal PP was stained with C6 during both late fetal developmental and postnatal stages. Neither the villous epithelial cell nor intestinal crypt basal cells were stained at any developmental stage. During the prenatal stages, FAE of the jejunal PP was C6-negative. However, a few C6-positive cells were distributed diffusely in some FAE of the jejunal PPs during the postnatal stages. The protein molecular weight of the antigen recognized by C6 was approximately 45 kDa. These data show that C6 is useful for identifying the FAE in ileal PPs and further suggest that differentiation of the FAE in these areas is independent of external antigens.
Animals ; Antibodies, Monoclonal/*immunology ; *Cattle ; Fetus ; Hybridomas ; Ileum/*ultrastructure ; Intestinal Mucosa/*immunology ; Peyer's Patches/*immunology/ultrastructure

In this review, we explain why and how rabbit monoclonal antibodies have become outstanding reagents for laboratory research and increasingly for diagnostic and therapeutic applications. Starting with the unique ontogeny of rabbit B cells that affords highly distinctive antibody repertoires rich in in vivo pruned binders of high diversity, affinity and specificity, we describe the generation of rabbit monoclonal antibodies by hybridoma technology, phage display and alternative methods, along with an account of successful humanization strategies.
Antibodies, Monoclonal* ; B-Lymphocytes ; Bacteriophages ; Humans ; Hybridomas ; Indicators and Reagents ; Sensitivity and Specificity

Monoclonal antibody producted by continuous perfusion culture was recovered and purified by expended bed adsorption chromatography. A packed bed column XK16/20 was used for method scouting with Streamline SP adsorbent. Two expended bed columns Streamline-25 and -50 were used for method optimization and pilot scale experiment, respectively. The recovery yield of monoclonal antibody was above 90% in a 5 - 7 fold enhanced purity and 10 fold increased concentration. According to the different concentration of monoclonal antibody in cell culture broth, about 18 - 50L fluid can be treated in a single cycle. MAb purification from lab scale (400mg per cycle) to a small pilot scale (2g per cycle) has been achieved. Compared with packed bed adsorption, the preparation cycle was half shortened, and the cost of production and the complexity of process were decreased markedly. It has been proven that a purification process based on expended bed adsorption technique is simple, efficient and economical.
Adsorption ; Antibodies, Monoclonal ; biosynthesis ; isolation & purification ; Bioreactors ; Cell Culture Techniques ; methods ; Chromatography ; methods ; Hybridomas ; cytology ; metabolism