1,2,3Assistant Professor, Kamalprakash Pharmacy College and Research Centre, Kherda, Tq. Karanja (Lad) Dist. Washim, Maharashtra-444107.
4Dr. Vithalrao Vikhe Patil Foundation's College of Pharmacy, Vilad ghat, Ahmednagar Maharashtra.
5,6,7,8Shri Sai institute of pharmacy and research Chhatrapati sambhajinagar, Maharashtra- 431006
The treatment of many illnesses, including as cancer, autoimmune conditions, and infectious diseases, has been completely transformed by monoclonal antibodies. Obstacles such immunogenicity, high production costs, restricted tissue penetration, and resistance development still exist despite their revolutionary effects. The methods of action, safety profiles, and clinical effectiveness of monoclonal antibodies are covered in this overview along with new developments and potential future paths. Advances in bio specific antibodies, antibody-drug conjugates, Nano bodies, checkpoint inhibitors, gene editing are emphasized as potential answers to existing challenges. Checkpoint inhibitors have revolutionized immunotherapy for cancer. The introduction of biosimilars has lowered prices and improved accessibility. These innovative advancements, their consequences for the treatment of illness, and potential avenues for further research are covered in this review. A detailed examination of industry trends, regulatory approvals, and clinical trial data is provided, offering insights into the changing MAB landscape. To find knowledge gaps and areas for improvement, new literature and expert comments are combined. Understanding the present situation and potential future developments of MAB treatment is aided by this review.
Monoclonal antibodies represent a groundbreaking advancement in biomedical science and therapeutic development. These highly specific proteins, created from identical copies (clones) of a single immune cell type, are engineered to precisely target specific antigens, making them indispensable tools in modern medicine. Since their inception in the 1970s, monoclonal antibodies have revolutionized the treatment of a wide array of diseases, including cancer, autoimmune disorders, and infectious diseases. Their journey, from discovery to widespread clinical application, is a testament to scientific innovation and their profound impact on patient care. The foundational technique for creating monoclonal antibodies was pioneered by Georges Köhler and César Milstein. Their method involves fusing a myeloma cell (a type of immortal cancer cell) with a specific B cell that produces the desired antibody. This fusion creates a hybridoma, a hybrid cell line capable of producing a uniform and specific antibody-referred to as a monoclonal antibody. Once isolated and purified, these antibodies can be utilized for therapeutic, diagnostic, or research purposes.
In recent years, monoclonal antibodies have become integral to the treatment landscape of various medical conditions. In oncology, they are employed for targeted therapies that specifically bind to cancer cells or associated proteins, minimizing damage to healthy tissues and enhancing therapeutic precision. In infectious diseases, they are used to neutralize pathogens or their toxins, offering a highly specific mechanism for combating infections. In immunology, monoclonal antibodies are designed to modulate immune responses, either by suppressing overactive immunity in autoimmune diseases or by enhancing it in conditions requiring immune activation. (1) Despite their transformative potential, the development and production of monoclonal antibodies present notable challenges. The process is highly complex, requiring advanced technology and expertise, which contributes to their high cost. Additionally, monoclonal antibody therapies can carry risks of side effects, including immune reactions and unintended interactions. Nevertheless, ongoing research and advancements in biotechnology are continually enhancing their efficacy, reducing production costs, and expanding their therapeutic applications.(2)
Monoclonal antibodies exemplify the intersection of science and medicine, offering a promising future for precision therapy and personalized medicine. As technology evolves, these powerful biological tools are poised to address an even broader spectrum of diseases, improving patient outcomes and transforming healthcare.
Development technique for monoclonal Antibodies:
Cell lines or clones derived from animals that have been vaccinated with the material under investigation produce these antibodies. Myeloma cells and B cells from the immunized animal are fused to create the cell lines (Köhler and Milstein 1975). One of two methods must be used to cultivate the cells in order to create the required mAb: either in vitro tissue culture or in vivo injection into the peritoneal cavity of a mouse that has been properly prepared (also known as mouse ascites). To get mAb with the necessary purity and concentration, additional processing of the tissue-culture supernatant and mouse ascetic fluid may be necessary. The mouse ascites technique is easily accessible, extensively known, and well-understood in many labs; however, the mice need to be closely observed in order to reduce any pain or discomfort brought on by an excessive build-up of fluid in the belly or by visceral invasion. If the in vitro tissue-culture method were as well-known and understood as the mouse ascites method, and if it produced the necessary amount of antibody with every cell line, it would be widely used. However, in vitro methods have been costly and time-consuming in comparison to the mouse ascites method, and they frequently failed to produce the necessary amount of antibody even with expert manipulation. The success rate has risen to over 90% thanks to modern in vitro techniques, which have also decreased expenses. (1-5)
Fig 2: Production of monoclonal antibodies by different techniques.
To initiate an immunological response, mice are first immunized with the desired antigens in the conventional mouse hybridoma approach. Myeloma cells and harvested splenocytes are united to create hybridoma cells, which manufacture antibodies continuously. Following screening, chimeric or humanized antibodies are produced using the chosen leads. (3)
produced from vaccinated or infected donors so that appropriate B lymphocytes can be isolated using flow cytometer. After RT-PCR, each B cell's VH and VL data are used to guide the production of human monoclonal antibodies (3-6)
The amount needed will depend on how the mAb is expected to be used (Marx and others 1997). Less than 0.1 g of mAb is needed for the majority of research undertakings and several analytical applications. Animal efficacy testing of novel mAb and the manufacturing of diagnostic kits and reagents both use medium-scale amounts (0.1–1 g). In this context, mAb production on a large scale is defined as more than 1 g. These greater amounts are employed for both therapeutic and standard diagnostic processes. For the identification of proteins, carbohydrates, and nucleic acids, monoclonal antibodies (mAb) have been and will remain crucial in biomedical research. Numerous chemicals that regulate cell division and replication have been clarified as a result of their utilization. Expanding our understanding of how molecular structure and function are related. Our knowledge of the host's reaction to infectious disease agents and the poisons they create, transplanted organs and tissues, spontaneously altered cells (tumors), and endogenous antigens (participated in autoimmunity) has improved as a result of these developments in basic biologic sciences. Furthermore, mAb's exceptional specificity makes it possible to diagnose and cure illnesses in both humans and animals. Since mAb-producing hybridomas can live forever in the right conditions, using fewer animals is linked to ongoing mAb production, particularly when in vitro techniques are used. "Animals chosen for the procedure should be of appropriate species and quality and the minimum number required to obtain valid results," according to the U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training (IRAC 1983). Techniques like computer simulation, mathematical models, and in vitro biological systems must to be taken into account. It is crucial to utilize animals properly, which includes avoiding or minimizing their pain, suffering, and distress when it is in line with good scientific principles. The excessive tumor burden in animals is expressly addressed in the Guide for the Care and Use of Laboratory Animals (NRC 1996, page 10), which notes that "sometimes, protocols contain methods that have not been previously encountered or that have the potential to result in uncontrollably severe pain or discomfort. The literature, veterinarians, researchers, and other experts on the effects in animals should be consulted for pertinent, unbiased information on the methods and the goal of the study. IACUCs must make sure that approved protocols adhere to the Public Health Service Policy on Humane Care and Use of Laboratory Animals (NIH 1996, page 7), which states that "procedures with animals... avoid or minimize discomfort, distress, and pain to animals (in a way that is) consistent with sound research design." Therefore, it is the scientist's responsibility to first think about using in vitro techniques to produce mAb. The researcher may ask for authorization to employ the mouse ascites approach if the in vitro generation of mAb is neither feasible nor reasonable. However, "IACUCs must determine that (I) the proposed use is scientifically justified, (ii) methods that avoid or minimize discomfort, distress, and pain (including in vitro methods) have been considered, and (iii) the latter [refers to in vitro methods] have been found unsuitable before they can approve proposals which include the mouse ascites method" (NIH 1997). The current committee was not charged with evaluating the procedures required to create a cell line that secretes antibodies.(7-12)
B-cells or plasma create antibodies, also known as immunoglobulin (Ig), which have a molecular weight of about 150 kDa. They are structurally composed of two functional components: a fragmented antigen- binding (Fab) region for target antigen recognition and Fc, or a crystallisable region linked to the effector mechanism. Two polypeptide chains— two light and two heavy-joined by disulphide bonds that provide stability and stiffness make up the antibody's two functional domains. Three constant domains (CH1, CH2, and CH3) and one variable domain (VH) make up the heavy chains. One constant domain (CL) and one variable domain (VL) make up the light chain. Together with VL and CL, VH and CH1 make up the Fab region, whereas CH2 and CH3 are the two segments that make up the Fc region. Additionally, post- translational changes in antibodies, like glycosylation in the Fc domain, stabilize and alter their ability to bind to Fc receptors. (2,13)
Types of monoclonal antibodies:
Mice are the complete source of these antibodies. Both the heavy and light chains' variable sections are derived from mice. The prefix "o-" or "-omab" is added to the general name to identify murine mAbs. For example, “rituximab” is a chimeric antibody that contains murine variable regions and is used to treat certain types of lymphoma and autoimmune diseases.(14)
Human constant regions and mouse variable regions make up chimeric antibodies. While the variable regions offer antigen specificity, the consistent sections dictate the antibody's effector actions. Chimeric monoclonal antibodies are identified by appending the suffix "- ximab" to the generic name. A chimeric antibody called "infliximab," for instance, targets tumor necrosis factor-alpha (TNF-?) and is used to treat inflammatory conditions including rheumatoid arthritis.
Only a minor amount of mouse sequence is included in the complementarity-determining regions (CDRs) of humanized antibodies, which primarily contain human sequence. Humanization lessens the immunogenicity of murine monoclonal antibodies. The suffix "- Zumba" is added to the generic name to create humanized mAbs. For instance, a humanized antibody called "trastuzumab" is used to treat breast cancer that is HER2-positive.(15)
Both in the constant and variable regions, fully human antibodies are made wholly from human sequences. They are made to be as compatible as possible with the human immune system while minimizing immunogenicity. The suffix "-umab" is added to the generic name to identify fully human mAbs. For instance, "adalimumab," a completely human antibody that targets TNF-?, is used to treat a number of autoimmune conditions, including psoriasis and rheumatoid arthritis.(16,17)
Mechanism of Action (MOA) of Monoclonal antibodies (mAbs):
Modern medicine relies heavily on monoclonal antibodies (mAbs), particularly in the fields of immunology, infectious diseases, and oncology. These modified antibodies work in a variety of ways and are made to target particular antigens, usually found on cells or pathogens. Their capacity to precisely engage and destroy targets or trigger immune effector actions to eradicate sick cells is the foundation of their therapeutic efficacy. Monoclonal antibodies have a complex mode of action (MOA), and depending on their target and design, different therapeutic antibodies use different processes.(18-26)
Antigen Neutralization:
In order to directly target pathogenic proteins or receptors implicated in disease processes, monoclonal antibodies are frequently employed. The antibody blocks the target antigen's biological function and stops it from interacting with other molecules by attaching itself to a particular epitope on the antigen.
Example:
Rituximab (Rituxan) depletes B lymphocytes by targeting their CD20. Since CD20 plays a role in B cell activation, blocking it can interfere with autoimmune processes or cancer, as is the case with diseases like rheumatoid arthritis and non-Hodgkin lymphoma. Targeting TNF-alpha, a cytokine that is essential in autoimmune conditions like rheumatoid arthritis and Crohn's disease, is infliximab (Regicide). Infliximab counteracts the pro-inflammatory effects of TNF-alpha by binding to it. (3)
Antibody-Dependent Cellular Cytotoxicity (ADCC)
Through the immune-mediated process of ADCC, the mAb attaches itself to its target on the surface of a tumor or infected cell. The antibody's Fc portion then engages with immune effector cells' Fc receptors, including neutrophils, macrophages, and Natural Killer (NK) cells. The effector cells are activated by this contact, which causes them to release cytotoxic chemicals (such granzymes and perforin) that cause the target cell to undergo apoptosis, or cell death.
Example:
Targeting the HER2 receptor, which is overexpressed in certain breast tumors, is Trastuzumab (Herceptin). Trastuzumab triggers ADCC, where immune cells eliminate HER2positive cancer cells, in addition to blocking HER2 signalling, which encourages tumor growth. Cetuximab is a monoclonal antibody that targets the epidermal growth factor receptor (EGFR). It is frequently used to treat head and neck tumors as well as colorectal cancer. Cetuximab-mediated ADCC has been linked to its anticancer properties. (4) (5)
Complement-Dependent Cytotoxicity (CDC)
Monoclonal antibodies have the ability to activate the complement system, a component of the innate immune response, in addition to ADCC. A mAb can start the traditional complement cascade when it attaches to its target antigen on the cell surface. A membrane attack complex (MAC), which is created as a result of this activation, damages the integrity of the cell membrane and causes cell death.
Example:
Rituximab: In individuals with non-Hodgkin lymphoma and other disorders involving CD20- positive B cells, rituximab not only induces ADCC but also activates CDC. Obinutuzumab: A glycoengineered anti-CD20 monoclonal antibody used to treat chronic lymphocytic leukaemia (CLL) that improves CDC in addition to ADCC. (6)
Inhibition of Signaling Pathways
Certain monoclonal antibodies are made to block particular signalling pathways that aid in the development of illness. In oncology, where tumor cells frequently depend on specific signalling pathways for growth, survival, and metastasis, this is especially pertinent.
Example:
Cetuximab (Erbitux): prevents the overexpression of EGFR (epidermal growth factor receptor), which is found in a number of malignancies, including colorectal and head and neck cancers. Cetuximab inhibits the signalling that EGFR activation increases cell growth and survival, which reduces the proliferation of tumors.
Pembrolizumab (Keytruda): Blocks the interaction between T cells' PD-1 receptor and tumor cells' PD-L1. Tumors can avoid immune detection because of this interaction, which often lowers the immune response. Pembrolizumab improves the immune response by inhibiting PD-1, which enables the body to combat cancer cells more effectively. (7)
Antibody-Drug Conjugates (ADCs)
ADCs, or antibody-drug conjugates, are a type of hybrid medicine that combines the powerful lethal effects of poisons or chemotherapy with the targeting power of monoclonal antibodies. ADCs provide targeted medication delivery to cancer cells by using a linker to connect a cytotoxicchemical to a monoclonal antibody. The conjugate is internalized and the cytotoxic drug is released inside the cell, causing cell death, once the mAb attaches to the target antigen.
Example:
Ado-Trastuzumab emtansine, also known as Kadcyla, is an ADC that combines the anti- HER2 Trastuzumab with DM1, a strong cytotoxic medication that prevents microtubule function. Breast cancer that is HER2-positive is treated with it. Adcetris or benuximab vedotin, is an ADC that targets CD30 and is used to treat malignancies that are CD30 positive, including Hodgkin lymphoma. Mon methyl auristatin E (MMAE), the cytotoxic medication, alters microtubule dynamics. (10)
Emerging monoclonal antibodies:
Recent studies have demonstrated the development of monoclonal antibodies for a number of illnesses, including infectious, autoimmune, and cancerous conditions, driven by breakthroughs in antibody engineering, combination treatments, and new mechanisms of action.(27-30)
Monoclonal antibodies for Various Diseases is as follows:
Rates of success of emerging monoclonal antibodies:
Table 1: Approval success rates for therapeutics monoclonal antibodies
|
Types of monoclonal antibodies |
Total number of MABS |
Number discontinued |
Number FDA approved |
Completion (%) |
Approval success (%) |
|
Humanized MABS, 1988-2006 |
131 |
53 |
11 |
49 |
17 |
|
Oncology humanized MABS |
62 |
22 |
4 |
42 |
15 |
|
Immunological humanized MABS |
45 |
19 |
5 |
53 |
21 |
|
Humanized MABS, 1988-1997 |
46 |
27 |
10 |
80 |
27 |
The comparatively high approval success rates of therapeutic mAbs-here defined as the probability that candidates pursuing clinical studies would ultimately obtain FDA approval—are a major factor in the present interest in these compounds. Compared to new chemical entities, chimeric and humanized mAbs often have greater success rates. Since humanized mAbs have been the subject of the most clinical research, they are the standard mAb type used to calculate success rates. The overall success rate for humanized mAbs that entered clinical studies between 1988 and 2006 was 17%, according to the data that is currently available (Table 1). The success rate of immunological mAbs was slightly higher (21%) than that of anticancer mAbs (15%) when the sample was stratified by therapeutic category. (2,31)
Table 2: Therapeutic monoclonal antibodies approved in United States: (2)
|
Generic name |
US trade name |
Therapeutic category |
|
Muromab-CD3 |
Orthoclone OKT3 |
Immunological |
|
Abciximab |
ReoPro |
Hemostasis |
|
Rituximab |
Rituxan |
Cancer |
|
Daclizumab |
Zenapax |
Immunological |
|
Basiliximab |
Simulect |
Immunological |
|
Palivizumab |
Synagis |
Anti-infective |
|
Infliximab |
Remicade |
Immunological |
|
Trastuzumab |
Herceptin |
Cancer |
|
Gemtuzumab ozogamicin |
Mylotarg |
Cancer |
|
Alemtuzumab |
Campath |
Cancer |
|
Ibritumomab tiuxetan |
Zevalin |
Cancer |
|
Adalimumab |
Humira |
Immunological |
|
Omalizumab |
Xolair |
Immunological |
|
Tositumomab-I131 |
Bexxar |
Cancer |
|
Efalizumab |
Raptiva |
Immunological |
|
Cetuximab |
Erbitux |
Cancer |
|
Bevacizumab |
Avastin |
Cancer |
|
Natalizumab |
Tysabri |
Immunological |
|
Ranibizumab |
Lucentis |
Opthalmic |
|
Panitumumab |
Vectibix |
Cancer |
CD, cluster of differentiation; (2)
Recent Advances in mAb Development:
Globally, unique antibody and protein synthesis services have undergone significant development and advancement since the 1960s. Scientists have created a variety of antibodies and their fragments with various architectures using potent techniques including phage display, B-cell amplification, and hybridoma technology with the aid of PCR. Due to their specificity and importance in the immune system's reaction to target antigens, monoclonal antibodies are essential. (12) Monoclonal antibodies (mAbs) have revolutionized the treatment of numerous diseases, particularly in the areas of oncology, autoimmune disorders, and infectious diseases. These biologics offer a very specialized tailored approach that reduces off-target effects because they are made to adhere to certain antigens. Recent advancements in monoclonal antibody development have improved patient outcomes, safety profiles, and efficacy while also increasing the therapeutic potential of these antibodies. This overview highlights some of the most important developments in the field of monoclonal antibodies during the past several years. (13,32)
The treatment of cancer has benefited greatly by the development of monoclonal antibody therapy. Nowadays, monoclonal antibodies are employed to modify immune checkpoint circuits or target antigens specific to tumors.
Additionally, monoclonal antibodies have become a potent therapy option for autoimmune illnesses, in which the body's tissues are mistakenly attacked by the immune system. Monoclonal antibodies can aid in the management of chronic inflammation and the prevention of tissue damage by specifically targeting immunological mediators or immune cells.
Clinical Efficacy and Safety:
Safety Concerns:
Regulatory Guidelines:
Safety Profile:
The following are typical adverse reactions (ARs) linked to monoclonal antibodies:
Challenge and future direction for the monoclonal antibodies:
In the treatment of many illnesses, especially cancer, autoimmune conditions, and infectious diseases, monoclonal antibodies (mAbs) have become essential. But using them presents a number of difficulties and issues for further research.
Future Directions:
CONCLUSIONS:
Although there are still issues, monoclonal antibodies have shown notable therapeutic benefit in a number of disorders. Promising answers to these problems can be found in emerging trends and technologies. Subsequent investigations ought to concentrate on refining the design of monoclonal antibodies, enhancing production procedures, and investigating combination treatments. The effectiveness and safety of monoclonal antibodies may be further improved by personalized medicine techniques that make use of gene editing and precision targeting. Monoclonal antibodies are positioned to continue to be a mainstay of contemporary medicine as the science develops, giving patients all over the world fresh hope.It is obvious that these targeted therapies will become more and more significant in the treatment of a variety of illnesses as the field of monoclonal antibodies develops. Monoclonal antibodies' noteworthy therapeutic advantages in a variety of clinical trials and real-world situations highlight their potential to improve patient outcomes. With new trends and technology providing creative answers to existing constraints, the development of monoclonal antibodies has a bright future. Researchers and doctors can fully realize the potential of these therapeutics by concentrating on improving production processes, improving antibody design, and investigating combination treatments. These tailored medicines will probably become more and more significant in the treatment of many ailments as a result of further research and development. In the end, monoclonal antibodies have a promising future and the ability to significantly improve the lives of millions of patients worldwide.
Conflicts Of Interests:
All authors have declared no conflict of interest.
REFERENCES
Anita Rathod*, Neha Khadse, Anjali Rathod, Ashok Chopane, Sapna Ghuge, Kavita Gaikwad, Manisha Khandagale, Priyanka Deshmukh, Emerging Monoclonal Antibodies for Treatment of Various Diseases: Recent Advances, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 1, 1309-1323. https://doi.org/10.5281/zenodo.14671974
10.5281/zenodo.14671974
