This is a working overview of recombinant-protein, written for readers who want more than a one-paragraph summary but less than a textbook.
Reviewed 2025-09-25. Anything still debated is marked as such rather than presented as settled.
There have been many efforts to engineer and improve new AAV variants for both clinical and research purposes. Such modifications include new tropisms to target specific tissues, and modified surface residues to evade detection by the immune system. Beyond opting for particular strains of recombinant AAV (rAAV) to target particular cells, researchers have also explored AAV pseudotyping, the practice of creating hybrids of certain AAV strains to approach an even more refined target. The hybrid is created by taking a capsid from one strain and the genome from another strain. For example:
In the European Union, a biological medicinal product is one of the active substance(s) produced from or extracted from a biological (living) system, and requires, in addition to physicochemical testing, biological testing for full characterisation. The characterisation of a biological medicinal product is a combination of testing the active substance and the final medicinal product together with the production process and its control. For example: Production process – it can be derived from biotechnology or from other technologies. It may be prepared using more conventional techniques as is the case for blood or plasma-derived products and a number of vaccines. Active substance – consisting of entire microorganisms, mammalian cells, nucleic acids, proteinaceous, or polysaccharide components originating from a microbial, animal, human, or plant source. Mode of action – therapeutic and immunological medicinal products, gene transfer materials, or cell therapy materials.
Affimer molecules, also known as adhiron, are small proteins that bind to target proteins with affinity in the nanomolar range. These engineered non-antibody binding proteins are designed to mimic the molecular recognition characteristics of monoclonal antibodies in different applications. These affinity reagents have been optimized to increase their stability, make them tolerant to a range of temperatures and pH, reduce their size, and to increase their expression in E.coli and mammalian cells.
Affimer molecules, also known as adhiron, are small proteins that bind to target proteins with affinity in the nanomolar range. These engineered non-antibody binding proteins are designed to mimic the molecular recognition characteristics of monoclonal antibodies in different applications. These affinity reagents have been optimized to increase their stability, make them tolerant to a range of temperatures and pH, reduce their size, and to increase their expression in E.coli and mammalian cells.
Genetically modified organism History of biotechnology An Asilomar Moment Archived 10 July 2013 at the Wayback Machine Original Asilomar Genetics Guidelines “Asilomar Conference.” Provides another summary of the Asilomar Conference. The Basics of Recombinant DNA provides an introduction to the science behind recombinant DNA. The Recombinant DNA Debate Archived 31 March 2007 at the Wayback Machine provides more details about the history of the debate surrounding the use of recombinant DNA technology. “Paul Berg: The 1980 Nobel Prize in Chemistry – Autobiography.” Provides an autobiography about Paul Berg. Science History Institute: The story of the 1975 Asilomar Conference on Recombinant DNA
Sources: en.wikipedia.org
This technology entails the joining of DNA from different species and the subsequent insertion of the hybrid DNA into a host cell. One of the first individuals to develop recombinant DNA technology was a biochemist at Stanford by the name of Paul Berg. In his experimental design in 1974, he cleaved (cut into fragments) the monkey virus SV40. He then cleaved the double helix of another virus, an antibacterial agent known as bacteriophage lambda. In the third step, he fastened DNA from the SV40 to DNA from the bacteriophage lambda. The final step involved placing the mutant genetic material into a laboratory strain of the E. coli bacterium. This last step, however, was not completed in the original experiment.
Boronate affinity chromatography consists of using boronic acid or boronates to elute and quantify amounts of glycoproteins. Clinical adaptations have applied this type of chromatography for use in determining long term assessment of diabetic patients through analysis of their glycated hemoglobin. Affinity purification of albumin and macroglobulin contamination is helpful in removing excess albumin and α2-macroglobulin contamination, when performing mass spectrometry. In affinity purification of serum albumin, the stationary used for collecting or attracting serum proteins can be Cibacron Blue-Sepharose. Then the serum proteins can be eluted from the adsorbent with a buffer containing thiocyanate (SCN−).
Andexanet alfa is a biologic agent, a recombinant modified version of human activated factor X (FXa). Andexanet alfa differs from native FXa due to the removal of a 34 residue fragment that contains the Gla domain. This modification reduces andexanet alfa's anticoagulant potential. Additionally, a serine to alanine (S419A) mutation in the active site eliminates its activity as a prothrombin to thrombin catalyst, but still allows the molecule to bind to FXa inhibitors. FXa inhibitors bind to andexanet alfa with the same affinity as to natural FXa. As a consequence, in the presence of andexanet alfa, natural FXa is partially freed, which can lead to effective hemostasis. In other words, it acts as a decoy receptor. Andexanet alfa reverses effect of all anticoagulants that act directly through FXa or by binding antithrombin III. The drug is not effective against factor IIa inhibitor dabigatran. Its activity is measured using the anti-Xa test, which is utilized to determine the amount of available factor Xa for coagulation
Affimer proteins were developed initially at the MRC Cancer Cell Unit in Cambridge then across two laboratories at the University of Leeds. Derived from the cysteine protease inhibitor family of cystatins, which function in nature as cysteine protease inhibitors, these 12–14 kDa proteins share the common tertiary structure of an alpha-helix lying on top of an anti-parallel beta-sheet. Affimer proteins display two peptide loops that can all be randomized to bind to desired target proteins, in a similar manner to monoclonal antibodies. Stabilization of the two peptides by the protein scaffold constrains the possible conformations that the peptides can take. This increases the binding affinity and specificity compared to libraries of free peptides, though can limit the target repertoire of Affimers.
AAV is of particular interest to gene therapists due to its apparent limited capacity to induce immune responses in humans, a factor which should positively influence vector transduction efficiency while reducing the risk of any immune-associated pathology. AAV is not considered to have any known role in disease. However, host immune system response and immune tolerance reduce the efficacy of AAV-mediated gene therapy. Host immune response has been shown to respond to the AAV vectors, the transduced cells, and the transduced proteins. The immune response can be subdivided into two categories: innate and adaptive, the latter of which is divided into humoral and cell-mediated.
Sources: en.wikipedia.org