A kodecyte (ko•de•cyte) is a

living cell The cell is the basic structural and functional unit of all forms of life. Every cell consists of cytoplasm enclosed within a membrane; many cells contain organelles, each with a specific function. The term comes from the Latin word meaning ' ...

that has been modified (koded) by the incorporation of one or more

function-spacer-lipid construct Function-Spacer-Lipid (FSL) Kode constructs (Kode Technology) are Amphiphile, amphiphatic, water wikt:dispersible, dispersible biosurface engineering constructs that can be used to engineer the surface of cell (biology), cells, viruses and organis ...

s (FSL constructs) to gain a new or novel biological, chemical or technological function. The cell is modified by the lipid tail of the FSL construct incorporating into the bilipid membrane of the cell. All kodecytes retain their normal

vitality Vitality (, , ) is the capacity to live, grow, or develop. Vitality is also the characteristic that distinguishes life, living from non-living things. To experience vitality is regarded as a basic psychological drive and, in philosophy, a comp ...

and functionality while gaining the new function of the inserted FSL constructs. The combination of dispersibility in

biocompatible Biocompatibility is related to the behavior of biomaterials in various contexts. The term refers to the ability of a material to perform with an appropriate host response in a specific situation. The ambiguity of the term reflects the ongoing ...

media, spontaneous incorporation into cell membranes, and apparent low toxicity, makes FSL constructs suitable as research tools and for the development of new diagnostic and

therapeutic A therapy or medical treatment is the attempted remediation of a health problem, usually following a medical diagnosis. Both words, ''treatment'' and ''therapy'', are often abbreviated tx, Tx, or Tx. As a rule, each therapy has indications an ...

applications.

The technology

Structural analogy of a sunflower to space filling models of selected FSL constructs

Kode FSL constructs consist of three components; a functional

moiety Moiety may refer to: __NOTOC__ Anthropology * Moiety (kinship), either of two groups into which a society is divided ** A division of society in the Iroquois societal structure in North America ** An Australian Aboriginal kinship group ** Native Ha ...

(F), a spacer (S) and a

lipid Lipids are a broad group of organic compounds which include fats, waxes, sterols, fat-soluble vitamins (such as vitamins A, D, E and K), monoglycerides, diglycerides, phospholipids, and others. The functions of lipids include storing ...

(L). Function groups on FSL constructs that can be used to create kodecytes include

saccharide A carbohydrate () is a biomolecule composed of carbon (C), hydrogen (H), and oxygen (O) atoms. The typical hydrogen-to-oxygen atomic ratio is 2:1, analogous to that of water, and is represented by the empirical formula (where ''m'' and ''n'' m ...

s (including

ABO blood group The ABO blood group system is used to denote the presence of one, both, or neither of the A and B antigens on erythrocytes (red blood cells). For human blood transfusions, it is the most important of the 47 different blood type (or group) cla ...

-related determinants,

sialic acid Sialic acids are a class of alpha-keto acid sugars with a nine-carbon backbone. The term "sialic acid" () was first introduced by Swedish biochemist Gunnar Blix in 1952. The most common member of this group is ''N''-acetylneuraminic acid ...

s, hyaluronin polysaccharides),

fluorophore A fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent chemical compound that can re-emit light upon light excitation. Fluorophores typically contain several combined aromatic groups, or planar or cyclic molecules with se ...

biotin Biotin (also known as vitamin B7 or vitamin H) is one of the B vitamins. It is involved in a wide range of metabolic processes, both in humans and in other organisms, primarily related to the utilization of fats, carbohydrates, and amino acids. ...

, and a range of

peptides Peptides are short chains of amino acids linked by peptide bonds. A polypeptide is a longer, continuous, unbranched peptide chain. Polypeptides that have a molecular mass of 10,000 Dalton (unit), Da or more are called proteins. Chains of fewer t ...

. Although kodecytes are created by modifying natural cells, they are different from natural cells. For example, FSL constructs, influenced by the composition of the lipid tail, are laterally mobile in the membrane and some FSL constructs may also cluster due to the characteristics of the functional group (F). As FSL constructs are anchored in the membrane via a lipid tail (L) it is believed they do not participate in

signal A signal is both the process and the result of transmission of data over some media accomplished by embedding some variation. Signals are important in multiple subject fields including signal processing, information theory and biology. In ...

transduction, but may be designed to act as agonists or antagonists of the initial binding event. FSL constructs will not actively pass through the plasma membrane but may enter the cell via membrane invagination and endocytosis. The "koding" of cells is stable (subject to the rate of turnover of the membrane components). FSL constructs will remain in the membrane of inactive cells (e.g. red blood cells) for the life of the cell provided it is stored in lipid free media. In the peripheral circulation FSL constructs are observed to be lost from red cell kodecytes at a rate of about 1% per hour. The initial "koding" dose and the minimum level required for detection determine how long the presence of "kodecytes" in the circulation can be monitored. For red blood "kodecytes" reliable monitoring of the presence of the "kodecytes" for up to 3 days post intravenous administration has been demonstrated in small mammals. The spacer (S) of a FSL construct has been selected so as to have negligible cross-reactivity with serum antibodies so kodecytes can be used with undiluted serum. By increasing the length of the FSL spacer from 1.9 to 7.2 nm it has been shown sensitivity can improve two-fold in red cell agglutination based kodecyte assays. However, increasing the size of the spacer further from 7.2 to 11.5 nm did not result in any further enhancement. Koded biomembrane

Technology Video

To view a simple video explaining how Kode Technology works, click the following link: https://www.youtube.com/watch?v=TIbjAl5KYpA

Methodology

Preparation of kodecytes and kodevirions

FSL constructs, when in solution (

saline Saline may refer to: Salt-related * Saline (medicine), a liquid with salt content to match the human body * Saline water, non-medicinal salt water * Saline, a historical term (especially American) for a salt works or saltern Places United States ...

) and in contact, will spontaneously incorporate into cell membranes. The methodology involves simply preparing a solution of FSL constructs in the range of 1–1000

μg In the metric system, a microgram or microgramme is a Physical unit, unit of mass equal to one millionth () of a gram. The unit symbol is μg according to the International System of Units (SI); the recommended symbol in the United States and Uni ...

/ mL, with the concentration used determining the amount of antigen present on the kodecyte. The ability to control antigen levels on the outside of a kodecyte has allowed for manufacture of quality control sensitivity systems and serologic teaching kits incorporating the entire range of serologic agglutination reactions. The actual concentration will depend on the construct and the quantity of construct required in the membrane. One part of FSL solution is added to one part of cells (up to 100%

suspension Suspension or suspended may refer to: Science and engineering * Car suspension * Cell suspension or suspension culture, in biology * Guarded suspension, a software design pattern in concurrent programming suspending a method call and the calling ...

) and they are incubated at a set temperature within the range of depending on temperature compatibility of the cells being modified. The higher the temperature, the faster the rate of FSL insertion into the membrane. For red blood cells incubation for 2 hours at 37 °C achieves >95% FSL insertion with at least 50% insertion being achieved within 20 minutes. In general, for carbohydrate based FSLs insertion into red blood cells, incubation for 4 hours at room temperature or 20 hours at 4 °C are similar to one hour at 37 °C. The resultant kodecytes do not required to be washed, however this option should be considered if an excess of FSL construct is used in the "koding process". Kodecytes can also be created ''

in vivo Studies that are ''in vivo'' (Latin for "within the living"; often not italicized in English) are those in which the effects of various biological entities are tested on whole, living organisms or cells, usually animals, including humans, an ...

'' by injection of constructs directly into the circulation. However this process will modify all cells in contact with the constructs and usually require significantly more construct than ''in vitro'' preparation, as FSL constructs will preferentially associate with free lipids. The ''in vivo'' creation of kodecytes is untargeted and FSL constructs will insert into all cells non-specifically, but may show a preference for some cell types. Diagnostic serological analyses including

flow cytometry Flow cytometry (FC) is a technique used to detect and measure the physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the ...

and scanning electron microscopy usually can't see a difference between "kodecytes" and unmodified cells. However, when compared with natural cells there does appear to be a difference between

IgM Immunoglobulin M (IgM) is the largest of several isotypes of antibodies (also known as immunoglobulin) that are produced by vertebrates. IgM is the first antibody to appear in the response to initial exposure to an antigen; causing it to also ...

and

IgG Immunoglobulin G (IgG) is a type of antibody. Representing approximately 75% of serum antibodies in humans, IgG is the most common type of antibody found in blood circulation. IgG molecules are created and released by plasma B cells. Each IgG ant ...

antibody reactivities when the functional group (F) is a monomeric peptide antigen. IgM antibodies appear to react poorly with kodecytes made with FSL peptides. Furthermore, FSL constructs may have a restricted antigen/epitope and may not react with a monoclonal antibody unless the FSL construct and monoclonal antibody are complementary. Kodecytes can be studied using standard histological techniques. Kodecytes can be fixed after "koding" subject to the functional moiety (F) of the FSL construct being compatible with the fixative. However, freeze cut or formalin-fixed freeze cut tissues are required because the lipid based FSL constructs (and other glycolipids) will be leached from the "kodecytes" in paraffin imbedded samples during the deparaffination steps.

Nomenclature

Koded membranes are described by the construct and the concentration of FSL (in

/ mL) used to create them. For example, kodecytes created with a 100 μg/mL solution of FSL-A would be termed A100 kodecytes. If multiple FSL constructs were used then the definition is expanded accordingly, e.g. A100+B300 kodecytes are created with a solution containing 100 μg/mL solution of FSL-A and 300 μg/mL solution of FSL-B. The "+" symbol is used to separate the construct mixes, e.g. A100+B300. If FSL concentrations are constant then the μg/mL component of the terminology can be dropped, e.g. A kodecytes. Alternatively unrelated constructs such as FSL-A and FSL-biotin will create A+biotin kodecytes, etc. If different cells are used in the same study then inclusion of the cell type into the name is recommended, e.g. RBC A100 kodecytes vs WBC A100 kodecytes, or platelet A100 kodecytes, etc.

Applications

Kode Technology has been used for the ''

in vitro ''In vitro'' (meaning ''in glass'', or ''in the glass'') Research, studies are performed with Cell (biology), cells or biological molecules outside their normal biological context. Colloquially called "test-tube experiments", these studies in ...

'' modification of murine

embryo An embryo ( ) is the initial stage of development for a multicellular organism. In organisms that reproduce sexually, embryonic development is the part of the life cycle that begins just after fertilization of the female egg cell by the male sp ...

spermatozoa A spermatozoon (; also spelled spermatozoön; : spermatozoa; ) is a motile sperm cell (biology), cell produced by male animals relying on internal fertilization. A spermatozoon is a moving form of the ploidy, haploid cell (biology), cell that is ...

zebra fish The zebrafish (''Danio rerio'') is a species of freshwater ray-finned fish belonging to the family Danionidae of the order Cypriniformes. Native to South Asia, it is a popular aquarium fish, frequently sold under the trade name zebra danio (and ...

epithelial Epithelium or epithelial tissue is a thin, continuous, protective layer of cells with little extracellular matrix. An example is the epidermis, the outermost layer of the skin. Epithelial ( mesothelial) tissues line the outer surfaces of man ...

endometrial The endometrium is the inner epithelium, epithelial layer, along with its mucous membrane, of the mammalian uterus. It has a basal layer and a functional layer: the basal layer contains stem cells which regenerate the functional layer. The funct ...

cells and

red blood cell Red blood cells (RBCs), referred to as erythrocytes (, with -''cyte'' translated as 'cell' in modern usage) in academia and medical publishing, also known as red cells, erythroid cells, and rarely haematids, are the most common type of blood cel ...

s to create cellular quality controls systems, serologic kits (teaching), rare

antigen In immunology, an antigen (Ag) is a molecule, moiety, foreign particulate matter, or an allergen, such as pollen, that can bind to a specific antibody or T-cell receptor. The presence of antigens in the body may trigger an immune response. ...

expression, add infectious markers onto cells, modified cell adhesion/interaction/separation/immobilisation, and labelling. It has also been

intravascular Blood vessels are the tubular structures of a circulatory system that transport blood throughout many animals’ bodies. Blood vessels transport blood cells, nutrients, and oxygen to most of the tissues of a body. They also take waste and c ...

ly infused for

modification of blood cells and neutralisation of circulating

antibodies An antibody (Ab) or immunoglobulin (Ig) is a large, Y-shaped protein belonging to the immunoglobulin superfamily which is used by the immune system to identify and neutralize antigens such as bacteria and viruses, including those that caus ...

and in ''in vivo'' imaging of circulating bone marrow kodecytes in zebrafish. Kode FSL constructs have also been applied to non-biological surfaces such as modified cellulose, paper, silica, polymers, natural fibers, glass and metals and has been shown to be ultra-fast in labelling these surfaces.

References

{{reflist, 30em, refs= Henry SM. Engineering the surface of red cells with synthetic glycolipids (KODETM CAE) to create ABO analytical sensitivity controls and xeno-modified cells. (invited lecture) 2nd International Symposium on ABO Incompatibility in Transplantation, Göteborg, Sweden, 2005 Xenotransplantation 2005; 12(5): 356 {{cite journal , last1 = Frame , first1 = Tom , last2 = Carroll , first2 = Tim , last3 = Korchagina , first3 = Elena , last4 = Bovin , first4 = Nicolai , last5 = Henry , first5 = Stephen , title = Synthetic glycolipid modification of red blood cell membranes , volume = 47 , issue = 5 , pages = 876–882 , year = 2007 , doi = 10.1111/j.1537-2995.2007.01204.x , journal = Transfusion , pmid = 17465953 , citeseerx = 10.1.1.494.2776 , s2cid = 18086433 {{cite journal , last1 = Hult , first1 = Annika K , last2 = Frame , first2 = Tim , last3 = Chesla , first3 = Scott , last4 = Henry , first4 = Stephen , last5 = Olsson , first5 = Martin L , title = Flow cytometry evaluation of red blood cells mimicking naturally-occurring ABO subgroups following modification with variable amounts of FSL-A and B constructs , volume = 52 , issue = 2 , pages = 247–251 , year = 2012 , doi = 10.1111/j.1537-2995.2011.03268.x , journal = Transfusion , pmid = 21812783 , s2cid = 5984970 {{cite journal , last1 = Flower , first1 = R , last2 = Lin P-H , first2 = Heathcote D , last3 = Chan , first3 = M , last4 = Teo , first4 = D , last5 = Selkirk , first5 = A , last6 = Shepherd , first6 = R , last7 = Henry , first7 = S , year = 2008 , title = Insertion of KODE peptide constructs into red cell membranes: Creating artificial variant MNS blood group antigens. ISBT Regional Congress, Macao SAR China, 2008". (P-396) , journal = Vox Sanguinis , volume = 95 , issue = Suppl 1, pages = 203–204 {{cite journal , last1 = Heathcote , first1 = D , last2 = Flower , first2 = R , last3 = Henry , first3 = S , year = 2008 , title = Development of novel alloantibody screening cells – the first example of the addition of peptide antigens to human red cells using KODE technology. ISBT Regional Congress, Macao SAR China, 2008". (P-303) , journal = Vox Sanguinis , volume = 95 , issue = Suppl 1, page = 174 {{cite journal , last1 = Henry , first1 = Stephen M , title = Modification of red blood cells for laboratory quality control use , volume = 16 , issue = 6 , pages = 467–472 , year = 2009 , doi = 10.1097/MOH.0b013e328331257e , journal = Current Opinion in Hematology , pmid = 19680123 , s2cid = 37416831 {{cite journal , last1 = Heathcote , first1 = Damien , last2 = Carrol , first2 = Tim , last3 = Wang , first3 = Jui-Jen , last4 = Flower , first4 = Robert , last5 = Rodionov , first5 = Igor , last6 = Tuzikov , first6 = Alexander , last7 = Bovin , first7 = Nicolai , last8 = Henry , first8 = Stephen , title = Novel antibody screening cells, MUT+Mur kodecytes, created by attaching peptides onto erythrocytes , volume = 50 , issue = 3 , pages = 635–641 , year = 2010 , doi = 10.1111/j.1537-2995.2009.02480.x , journal = Transfusion , pmid = 19912581 , s2cid = 20952307 {{cite journal , last1 = Chesla , first1 = S , last2 = Henry , first2 = S , last3 = Eatz , first3 = R , last4 = Sinor , first4 = L , title = Solid phase syphilis test utilizing KODE technology , volume = 50 , pages = 196A–197A , year = 2010 , doi = 10.1111/j.1537-2995.2010.02833_1.x , pmid = 20815863 , journal = Transfusion , s2cid = 222195124 {{cite journal, url=http://carbohyd.siobc.ras.ru/printable.php?id=381, vauthors=Komarraju S, Chesla S, Bovin N, Henry S , title= Syphilis-kodecytes – novel function-spacer-lipid (FSL) modified red cells capable of sensitive and specific detection of syphilis antibodies, year=2010, journal= FEBS Journal , volume=277 , issue= S1, pages=97–98, doi=10.1111/j.1742-4658.2010.07680.x, pmc=7164047 , hdl=10292/2142 , hdl-access=free {{cite journal , vauthors=Blake D, Lan A, Love D, Bovin N, Henry S , title=Fluorophore-kodecytes – fluorescent function-spacer-lipid (FSL) modified cells for in vitro and in vivo analyses, doi=10.1111/j.1742-4658.2010.07680.x , journal=FEBS Journal , issue=1 , year=2010 , pages=37–271 , volume=277 , pmc=7164047 , hdl=10292/2142, hdl-access=free {{cite journal , last1 = Nadarajan , first1 = V.S. , last2 = Laing , first2 = A. A. , last3 = Saad , first3 = S. M. , last4 = Usin , first4 = M , title = Prevalence and specificity of red-blood-cell antibodies in a multiethnic South and East Asian patient population and influence of using novel MUT+Mur+ kodecytes on its detection , volume = 102 , issue = 1 , pages = 65–71 , year = 2011 , doi = 10.1111/j.1423-0410.2011.01507.x , journal = Vox Sanguinis , pmid = 21592136 , s2cid = 20297050 , doi-access = free {{cite journal , last1 = Henry , first1 = Stephen , last2 = Komarraju , first2 = Sarvani , last3 = Heathcote , first3 = Damien , last4 = Rodinov , first4 = Igor L , title = Designing peptide-based FSL constructs to create Miltenberger kodecytes , volume = 6 , issue = 2 , pages = 306–312 , year = 2011 , doi = 10.1111/j.1751-2824.2011.01505.x , journal = ISBT Science Series , s2cid = 82441272 , doi-access = free {{cite journal , last1 = Georgakopoulos , first1 = T , last2 = Komarraju , first2 = Sarvani , last3 = Henry , first3 = Stephen , last4 = Bertolini , first4 = Joseph , title = An improved Fc function assay utilising CMV antigen coated red blood cells generated with synthetic Function-Spacer-Lipid constructs , volume = 102 , issue = 1 , pages = 72–78 , year = 2011 , doi = 10.1111/j.1423-0410.2011.01512.x , journal = Vox Sanguinis , pmid = 21749406 , s2cid = 9758322 {{cite journal , last1 = Oliver , first1 = Caroline , last2 = Blake , first2 = Debbie , last3 = Henry , first3 = Stephen , title = In vivo neutralization of anti-A and successful transfusion of A antigen incompatible red cells in an animal model , volume = 51 , issue = 12 , pages = 2664–2675 , year = 2011 , doi = 10.1111/j.1537-2995.2011.03184.x , journal = Transfusion , pmid = 21599675 , s2cid = 205724219 {{cite journal , last1 = Oliver , first1 = Caroline , last2 = Blake , first2 = Debbie , last3 = Henry , first3 = Stephen , title = Modeling transfusion reactions and predicting in vivo cell survival with kodecytes , volume = 51 , issue = 8 , pages = 1723–1730 , year = 2011 , doi = 10.1111/j.1537-2995.2010.03034.x , journal = Transfusion , pmid = 21303367 , s2cid = 24736518 {{cite journal , last1 = Blake , first1 = Debbie A , last2 = Bovin , first2 = Nicolai V , last3 = Bess , first3 = Dan , last4 = Henry , first4 = Stephen M , title = FSL Constructs: A Simple Method for Modifying Cell/Virion Surfaces with a Range of Biological Markers Without Affecting their Viability , volume = 54 , issue = e3289 , year = 2011 , doi = 10.3791/3289 , pmid = 21847082 , pmc = 3211133 , journal = Journal of Visualized Experiments {{cite journal , last1 = Lan , first1 = C-C , last2 = Blake , first2 = D , last3 = Henry , first3 = S , last4 = Love , first4 = D R , title = Fluorescent Function-Spacer-Lipid construct labelling allows for real-time in vivo imaging of cell migration and behaviour in zebrafish (Danio rerio) , year = 2012 , doi = 10.1007/s10895-012-1043-3 , journal = Journal of Fluorescence , pmid=22434405 , volume=22 , issue=4 , pages=1055–63 , s2cid = 14406691 , hdl = 10292/3475 , hdl-access = free {{cite journal , last1 = Korchagina , first1 = Elena , last2 = Tuzikov , first2 = Alexander , last3 = Formanovsky , first3 = Andrey , last4 = Popova , first4 = Inna , last5 = Henry , first5 = Stephen , last6 = Bovin , first6 = Nicolai , title = Toward creating cell membrane glycolandscapes with glycan lipid constructs , year = 2012 , doi = 10.1016/j.carres.2012.03.044 , journal = Carbohydrate Research , pmid=22551471 , volume=356 , pages=238–46 {{cite book , last1 = Henry , first1 = Stephen , last2 = Rodionov , first2 = Igor , title = FSL-RFG(Maleimide) FSL Construction Kit Technical Bulletin , publisher = Scholarly Commons , year = 2012 , hdl = 10292/2241 {{cite book , last1 = Henry , first1 = Stephen , last2 = Perry , first2 = Holly , title = FSL-A+B(tri) Serologic Teaching Kit Technical Bulletin , publisher = Scholarly Commons , year = 2012 , hdl = 10292/2827 {{cite journal , last1 = Henry , first1 = Stephen , last2 = Barr , first2 = Katie , last3 = Oliver , first3 = Caroline , title = Modeling transfusion reactions with kodecytes and enabling ABO-incompatible transfusion with function-spacer-lipid constructs , journal = ISBT Science Series , date = 2012 , volume = 7 , issue=1 , pages = 106–111 , doi = 10.1111/j.1751-2824.2012.01563.x

External links

Kodeycte.com

How Kode Technology works

Applications of kodecytes

CSL Limited, CSL application of kodecytes Biochemistry Biotechnology Laboratory techniques Molecular biology techniques Protein methods Nanotechnology