One of the goals of protein genetic engineering is the stabilisation of proteins,
see J.T. Kellis et al., "Protein stabilisation by engineered metal chelation", Biotechnology 9
: 994-5. They inserted a metal-chelating site (His-X3-His) into a cytochrome c, and the protein was stabilised. On engineered antibodies
(Sept 1991), 128. On metabolic engineering see PNAS
88: 8784-6; Applied & Env. Microbiology
A general comment on characterising recombinant proteins is in Biotechnology 9 : 921-4. On laser manipulation of atoms and particles see Science 253; 861-6.
Comments on biosensors and the use of polymers in their manufacture is Science 253: 1097-8. On biomimicry see Science 253: 966-8, & SA (Sept 1991), 122-5.
Targeting of recombinant antibodies to the surface of E. coli
is described in Biotechnology
9: 1369-72. Such targeting may also be useful in the production of live vaccines.
See also B.K. Klein et al., "Secretion of active bovine somatotropin in E.coli
On yeast systems for recombinant DNA encoded protein production see Biotechnology 9: 1067-72. On xylitol production by yeast see Biotechnology 9: 1090-5. On the search for new compounds see P.D. Crittenden & N. Porter, "Lichen-forming fungi: potential sources of novel metabolites", TIBTECH 9: 409-14. These fungi have not been examined by pharmacologists.
An economic analysis of the future use of biomass in Europe is J. Swinnen & E. Tollens, "Bulk chemicals from biomass in the EC: Feasibility and potential outlets", Bioresource Technology 36: 277-91. With the present techniques, they conclude that biomass will only play a significant role if subsidies were introduced, or the price of oil became higher. On ethanol production from mixed bacterial cultures see Biotechnology Letters 13: 761-4. On the future impact of industrial lipases see TIBTECH 9: 360-3.
The use of reverse sample genome probing for detection of bacteria in oil is described in Applied & Environmental Microbiology 57: 3070-8.
On the use of mammalian cell culture in bioreactors see Biotechnology 9: 805-12. The production of TPA is described, as well as a general review. On the use of myeloma based gene expression systems for the production of large mammalian proteins see TIBTECH 9: 109-15. On mixed animal cell cultures to produce recombinant products see TIBTECH 9: 406-7. On the use of insect cells to produce human amyloid b-protein see PNAS 88: 10307-11.
On antibody engineering see; M.W. Fanger & P.M. Gurye, "Bispecific antibodies for
targeted cellular cytotoxicity", TIBTECH
9: 375-80. These antibodies can have specificity for both a drug and a tumor cell,
so that drugs concentrate at the tumor cells. On the use of molecules for cancer
treatment see I. Pastan & D. Fitzgerald, "Recombinant toxins for cancer treatment",
254: 1173-7. Such toxins can be developed so that they are selective, and fusions
of toxin with growth factor or antibodies are already made.
On the research on the production of antibodies in plants see TIBTECH 9: 107-8. On the cloning of antibodies in bacteria see TIBTECH 9: 169-75. Inorganic molecules have been synthesized that mimic the hypervariable regions of antibodies, see GEN (Nov/Dec 1991), 43. They may be more useful than some synthetic peptides, and could be made to mimic any sequence of amino acids, provided that their conformation is correct.
On obtaining multienzyme systems by gene fusions see TIBTECH 9: 226-31. On protein engineering see the journal Protein Engineering 4: 709-10. On enzyme engineering for use in nonaqueous solvents see Biotechnology 9: 1073-7; TIBTECH 9: 394-8. This may allow different reactions and reaction rates, using organic solvents. On cofactor engineering see TIBTECH 9: 343-6. On stabilising of proteins see TIBTECH 9: 259-60. On a chimeric tissue plasminogen activator (TPA) with high affinity for fibrin see PNAS 88: 10337-10341. Attempts to redesign TPA for improved therapy will need clinical tests to prove the efficacy, and safety; TIBTECH 9: 86-90.
Biocomputers which use enzymes as flip flops for signals are discussed in NS (7 Dec 1991), 23; Science 254: 1308-10. On improving the rate of energy capture from light over photosynthesis see Nature 353: 698-9. A review on biosensors is Enzyme Microb. Technology 13: 946-55; see also TIBTECH 9: 260-1.
There is an effort in Japan to obtain more microorganisms from the ocean depths, there
is a new research program including the world's deepest diving manned submarine and
a base research facility to culture such microbes under very high pressure and over
a range of temperatures from below freezing to above boiling; Science
255: 28. On the use of low temperature microbes for high yields see a short discussion
of the use of microbes isolated from Antarctica in Biotechnology
10: 7. On microorganism repositories see Biotechnology
As follows of the newspapers will be aware, there were also some organisms grown on a recent space shuttle flight. The results of some recent data on studies performed on the more permanent Soviet space laboratory are in Biotechnology 10: 128; J. General Microbiology 137: 2839.
Some other recent papers on fermentation and cell culture to produce useful products are; production of Von Willebrand factor, Biotechnology 10: 66-73; on the production of recombinant antibody from myeloma cells, Biotechnology 10: 169-75. The use of Whey permeate, which is a waste product of cheese production, as a medium for the growth of yeast see Biotechnology 10: 82-5. This approach could solve the waste problem, as well as utilise it in a useful way. On the production of surface-active compounds from microorganisms see a review in Biotechnology 10: 60-5. On large scale cell culture see Biotechnology 10: 31-2.
A review of antigenized antibodies is in Nature
355: 476-7. Oligopeptides are expressed in the hypervariable loops of antibodies
and these can modify the binding of the antibodies. On the expression of a humanized
antibody fragment in E. coli
10: 163-7. The production of large quantities is necessary for these antibodies to
be used for therapy. On antibody assisted protein folding see Biotechnology
There are increasing numbers of biosensors being developed, and some are listed in Biotechnology 10: 129. On the use of biomimetrics see Science 255: 284-6, for a short review titled "animating the material world".
The production of human growth hormone in bacteria has been troubled by the misfolding of the protein in microbes which leads to the formation of aggregates. A method to overcome this using the simultaneous overproduction of a chaperonin molecule, which aids protein folding, is P. Blum et al., "Dna-K-mediated alterations in human growth hormone protein inclusion bodies", Biotechnology 10: 301-4. The production of the related human protein, growth hormone-releasing hormone in E.coli is described in Biotechnology 10: 315-9.
The use of
, and the construction of a diverse human antibody library in phage, is commented
on in BMJ
304: 585-6. Clinical trials of one such antibody, Campath-1H, against non-Hodgkin's
lymphoma have begin in the USA and will begin in Europe; NS
(29 Feb 1991), 18.
The technique of site-directed mutagenesis has been used to create mutants of genes, to produce proteins where particular amino acids are changed, which may alter properties. By optimizing the nucleotide mixtures in such reactions, the resultant amino acid codon becomes less random, called semi-random mutagenesis; Biotechnology 10: 297-300.
Other topics of interest include; biomimetics in materials research, Science 255: 1098-1105; 684; ribozymes and industry, Biotechnology 10: 256-62.
A review of methods to develop drugs from traditional medicinal plants is in Chemistry & Industry
(20 April 1992), 290-3. A review on the development of therapeutic molecules based on
oligonucleotides is in TIBTECH
10: 87-91. A review on therapeutics based on triple helix technology is in TIBTECH
10: 132-6 (triple helix means oligonucleotides that can bind to the double helix
of DNA, forming a triple helix). Computerised drug design is discussed in Science
256: 440-2. Papers on the subject of RNA catalysis, including newly found roles
where RNA is involved in catalysis are: Science
256: 1396-7, 1402-4, 1416-23.
An article on a class of small molecules which act by cleaving DNA and which may have therapeutic roles is K.C. Nicolau et al., "Designed enediynes: a new class of DNA-cleaving molecules with potent and selective anticancer activity", Science 256: 1172-8. On research on combating the common cold, see S.M. Edgington, "Bauhaus biology: biotechnology and the common cold", Biotechnology 10: 502-7.
A discussion of bacteriophage screening systems for antibody production is in D.J. Chiswell & J. McCafferty, "Phage antibodies: will new 'coliclonal' antibodies replace monoclonal antibodies?", TIBTECH 10: 80-84; see also p.75-6. A conference report on humanised antibody engineering is in TIBTECH 10: 112-3. Anti-endotoxin antibodies to treat sepsis are another possible use of antibody treatment; NEJM 326: 1151-7. The use of antibodies in enzyme reactions is illustrated by J.R. Jacobsen et al., "An efficient antibody-catalysed aminoacylation reaction", Science 256: 365-7. Protein analysis and design is discussed in TIBTECH 10: 141-3, and the knowledge gained from the human genome project should greatly increase the repertoire of designs. On new products for design of proteins see Nature 356: 545-6. The genetic code has been expanded to add a non standard amino acid, which could also add diversity to protein design; Nature 356: 537-9.
A review on the issue of drug delivery is J. Kost & R. Langer, "Responsive polymer systems for controlled delivery of therapeutics", TIBTECH 10: 127-31.
A technique under license to a US company Exogene involves expression of hemoglobin
genes in the cells of yeast, which increases the oxygen capacity of the cells and
thus increases the production of other recombinant proteins; GEN
12(11), 1, 19. In bacteria increased production can be obtained by plasmids which
over-replicate (to 1000 copies per genome), K. Nordstrom & B.E. Uhlin, "Runaway-replication
plasmids as tools to produce large quantities of proteins from cloned genes in bacteria", Biotechnology
10: 661-6. It is possible to make the protein encoded by the plasmid as 10-50% of
the total protein.
An example of metabolic engineering of yeast is S. Picataggio et al., "Metabolic engineering of Candida tropicalis for the production of long-chain dicarboxylic acids", Biotechnology 10: 894-8. The produced acids can be used for industrial use.
A review on the production of flavours by microorganisms is in Process Biochemistry 27: 195-215. A review (in German) of the genus Chlorella , an algae used in biotechnology, is in Naturwissenschaften 79: 260-5.
It has proved possible to add a metal-binding site to the enzyme trypsin, so that the enzymatic activity is dependent upon the addition of zinc, nickel or copper; GEN 12(8), 1, 23. The addition of a chelating agent like EDTA can then turn off the enzyme. See also PNAS 89: 6803-7 for enzyme engineering.
A review of research on producing
in bioreactors is S.M. Edgington, "New horizons for stem-cell bioreactors", Biotechnology
10: 1099-1106. It looks at the company SyStemix, which has received a patent for
the human hematopoietic stem cell.
The production of the enzyme streptokinase in E.coli as 25% of total protein, is reported in Bio 10: 1138-42. It offers a good source of the protein, which is already used in medicine. A method for the production of a chimeric human antibody in Chinese Hamster Ovary Cells is described in Biotechnology 10: 11221-7.
With the aim of making a targeted 'magic bullet' to destroy disease-promoting cells, bacterial toxins fused to cytokine-receptor binding regions have been made by genetic engineering by a company Seragen in the USA; GEN (1 Sept 1992), 22-3. They are claimed to be better than targetted molecules based on monoclonal antibodies, which is the method being explored at the moment by many companies. The use of cancer vaccines is discussed in an article in Newsweek (19 Oct 1992), 49-50.
Research on anti-idiotypic monoclonal antibodies is discussed in Biotechnology 10: 950. A review on protein engineering is D. Medynski, "Genetic approaches to protein structure and function: point mutations as modifiers of protein function", Biotechnology 10: 1002--6. Modifying function of proteins by glycosylation is reported in Biotechnology 10: 1143-7. A review of screening for pharmaceuticals based on binding to receptor proteins is in Biotechnology 10: 973-80. The use of molecular biology could provide cleaner systems for screening for possible new drugs than current methods, often using animal cells, tissues or animals. The development of drugs is discuused in a review I.D. Kuntz, "Structure-based strategies for drug design and discovery", Science 257: 1078-82.
Methods that are being investigated to increase the half-life of biopharmaceuticals are discussed in GEN (Aug 1992), 1, 14. One method is to make a conjugate with larger proteins to increase the circulation life, using site-directed mutagenesis, or altering glycosylation to make it less immunogenic so the body's immune system does not destroy it so rapidly.
Enzyme stability in vitro can be improved by drying in the presence of a sugar trehalose; Biotechnology 10: 1007-11. A method to immobilise biomolecules on 'inert' surfaces using trifluoromethyl-aryl-diazirines to serve as linkers is described in Biotechnology 10: 1026-8.
A clinical trial of using an antisense oligodeoxynucleotide to turn off the expression of the product of the p53 cancer-related gene has been undertaken; GEN (Aug 1992), 1, 28. The use of DNA and RNA as therapeutics is reviewed in Bio 10: 993-6.
On biosensors for NO on a cellular size see Nature 358: 623, 676-8. NO is important as a messenger for killing of tumour cells and bacteria by macrophages, and acting as a neurotransmitter. In 1990 a patent application (#491059) was filed in Europe, which has just been published, for a method to crystalise DNA in a precise manner on flat surfaces; NS (17 Oct 1992), 18. This may be useful for production of chips for electrical uses. On angiogen and drugs see Bio 10: 981-5. The progress in computer models of life and ecology is reported in Science 257: 1040--2.
Cd-metal binding chimeric antibodies have been expressed in E.coli; PNAS
89: 9754-8. The use of an antibody as a catalyst to rearrange a peptide bond is
described in Science
258: 803-5. "Primatization" of recombinant antibodies for therapeutic use is described
10: 1455-60. A chimeric macaque/human antibody against human CD4 is being used
as a possible therapy for arthritis. A review on adaption of antibodies for clinical
use is in BMJ
305: 1348-52, and a history of monoclonal antibodies BMJ
305:1269-72. The production of antibodies without immunisation, but using bacteria
and bacteriophages, is reviewed in Science
An editorial on current research in protein engineering by J. Hodgson is in Biotechnology 10: 1433-4. Design of proteins and optimisation by experiments may produce more efficient enzymes and molecules for human application than has been possible in nature. The use of computers in biology, so-called artificial life, is the topic of a book review in Nature 360: 25-6. The use of peptoids (oligomers of N-substituted glycines) for drug discovery is proposed as a good way to develop drugs in PNAS 89: 9367-71. The use of artificial intelligence in drug design is also growing; SA (Nov 1992), 87. A new technology to use laser beams to manipulate the position of molecules has been developed; Nature 360: 493-5.
The technology of generating and screening peptides using genetic engineering for
drug design is discussed in GEN
(Dec 92), 1, 22-3. Recent research use of drugs such as cocaine to study brain function
is reviewed in Science
258 (1992), 1882-4. Engineering using ribozymes can use natural selection; Science
258 (1992), 1910-5; Nature
361: 119-20, 182-5. On the general use of directed evolution to select molecules
(Dec 1992), 48-55.
The possibilities for design improvement of naturally-occuring human hormones and cytokines to treat disease is discussed in Nature 360 (1992), 711-2. The molecule tumour necrosis factor has some bad side effects, so mutants that have less side effects may be more useful, and a review of strategies is; Nature 361: 206-7. The general topic and its relevance to drug discovery is reviewed, with many examples, in Biotechnology 10 (1992), 1529-4.
Method for enhancing the production of monoclonal antibodies, by inoculating human spleen cells, and expression of the selected antibody genes in M13 bacteriophage, are reported in J. Immunology 149 (1992), 3903-13, 3914-20; GEN (15 Jan 1993), 1, 19. A polymer coating around antibodies can increase their in vivo life; NS (12 Dec 1992), 20.
Catalytic antibodies are being developed for various applications; Science 259: 469-70. Reviews of antibody therapy for cancer are JAMA 269: 78-81; BMJ 305 (1992), 1424-9; Science 259: 310-11; Biotechnology 11: 156-7, 159-62.
A review of some commercially available biosensors is in Nature 361: 186-7. A review of methods to increase production of recombinant proteins in E.coli is Biotechnology 10 (1992), 1550-6. A special issue of The FASEB Journal 7 (1): 1-239, features many reviews on RNA.
A comparison of the costs of using animal cell culture or bacteria for production of TPA is in Biotechnology 11: 349-57. A paper reporting the hydrolysis of cellulose by E.coli expressing cellulase enzyme is Biotechnology 11: 491-5. A book review on Applied Molecular Genetics of Filamentous Fungi is in TIBTECH 11: 69.
A special issue of J. Biotechnology 28(1), 1-138 is on the industrial impacts of protein engineering . A general introduction to the issue and topic is on p. 1-23. A report from the Miami Biotechnology symposium on protein engineering is in Biotechnology 11: 278-9.
A review of techniques for engineering antibodies is J. Singh Sandhu, "Protein engineering of antibodies", CRC Critical Reviews in Biotechnology 12 (1992), 437-62. Genetic methods to generate antibodies are reported in Biotechnology 11: 503-7; PNAS 90: 1160-5. A 61 amino acid antibody, called a "minibody" has been engineered based on the variable region of immunoglobulin, Nature 362: 293-4, 367-9. General regulatory and clinical issues of monoclonal and therapeutic antibodies are discussed in TIBTECH 11: 40-4. An animal model trial is H. Takahashi et al., "Inhibition of human colon cancer growth by antibody-directed human LAK cells in SCID mice", Science 259: 1460-3. A general review is T. Boon, "Teaching the immune system to fight cancer", SA (March 1992), 32-9.
Papers on the use of computers in development of drugs are in Biotechnology 11: 285-9, 465-8, 472-3. The use of naturally occuring drugs isolated from organisms is the traditional way of finding new drugs, and still is useful, SA (March 1992), 101-2. On the use of mimics of biological designs for industry see Time (8 March 1992), 40-1; Science 259: 1282-7; NS (13 Feb 1993), Supplement 12-16. An artificial enzyme, synthesised from chemicals, is described in NS (3 April 1993), 16; Science 259: 1675, 1699.
A review of adoptive
to treat cancer is in SA
(Medicine-special edition 93), 94-101. A review of immune intervention in general
is in Science
260: 937-44. It discusses the developments that could result in molecular vaccination
and control of graft rejection in tissue transplants. New techniques for conjugating
cytotoxic agents to antibodies are reviewed in GEN
(15 March 1993), 6, 20. A review of research to develop antibody therapies to treat
is in JAMA
Naturally occuring antibodies have been found without light chains; Nature 363: 446-8. This means that further possibilities for protein engineering are opened. On generation of antibody diversity in VDJ recombination see Nature 363: 625-7. Colorimetric antibodies for assays have been made by combining two genes; Biotechnology 11: 601-5. The use of a filamentous fungi to produce antibodies by engineering is reported in Biotechnology 11: 591-5. Also on antibody production and clonal selection in the immune system see Nature 363: 208, 271-3. Chemical reactivity can also be controlled by antibodies; Science 260: 337-9, and we can expect industry to use the selectivity that enzymes provide in the future much more.
Protein engineering of tumor necrosis factor is reviewed in GEN (15 March 1993), 1, 16. TNF is toxic, and reduced toxicity will enable its more effective use in cancer therapy. A review on the physiological roles of TNF in wasting and obesity is in Cell 73: 625-7, see also Cell 73: 213-6, 431-45, 447-56. A new family of cytokines with homology to TNF has been identified.
The use of complex oligosaccharides for biological signaling is reviewed in GEN (15 April 1993),10-1. A report from the US Association of Pharmaceutical Scientists looks at new ways for delivering drugs in vivo; GEN (15 Feb 1993), 1, 10, 25; see also Science 260: 912-3. Polypeptides can be released in a controlled manner by coupling to polyanhdrides, PNAS 90: 4176-80. The development of liposomes has been rapid and their commerical use in the USA is expected soon; GEN (15 Feb 1993), 6-7.
Drugs specially designed to attack the influenca virus are reported to inhibit infection in animal models; Nature 363: 401-2, 418-23. On general drug design using molecular biology see Science 260: 910-2, 918-9; NS (29 May 1993), 35-8. The mechanism of recombination between virus-associated RNAs is reported in Science 260: 801-5. A 4-stranded nucleic acid, tetrad, is reported in Nature 363: 499-500. On the use of oligonucleotides as therapy see Science 260: 1510-3.
The use of biosensors to assess the health of organisms in the environment is the topic of a new book, Animal Biomarkers as Pollution Indicators, David Peakall, reviewed in EST 27: 1003.
A review of biocatalysis and immobilised bioreactors is in Biotechnology 11: 690-5; and on bioprocess engineering, Enzyme & Microbial Technology 15: 541-3. The genetic modification of yeast to produce more alcohol from starch by inclusion of rice amylase gene is reported in Biotechnology 11: 606-10.
A Texas company Cytoclonal Pharmaceuticals has been chosen to have the licensing rights
by Montana State University to the technique for production of
from a fungus; GEN
(July 1993), 1, 16; Science
261: 539. A paper reporting the development of protaxols, compounds that will increase
the effectiveness of taxol therapy in the body is in Nature
More on drug design in GEN (July 1993), 1, 36. The use of the space shuttle as a vehicle for space experiments is discussed in GEN (July 1993), 1, 10. A review on biosensors is in TIBTECH 11: 122-30; see also Science 260: 1711. A review on the use of metal compounds in therapy is in Science 261: 699-700, 725-30; and on ribozymes, on pp. 709-14. On antisense techniques see Science 260: 1510-3.
High activity mammalian cytochrome P-450 has been expressed in yeast; TIBTECH 11: 131-5; which may allow detoxification of drugs, and other uses. A conference review of the 1993 Miami Biotech Winter Symposium on protein engineering is in TIBTECH 11: 111-4. A review on engineering proteins for nonnatural environments is FASEB J 7: 744-9.
The use of monoclonal antibodies for imaging and therapy is reviewed in GEN (Aug 1993), 6-7, 10. A cure for human cancers that were xenografted into mice by antibody conjugates is in Science 261: 212-5. A review of catalytic antibodies is in Hospital Practice (15 July 1993), 53-9. Humanised antibody research is rapidly expanding, SA (July 1993), 85-7; and on mice with human antibodies, NS (21 Aug 1993), 20.
Immunotherapy using recombinant
is discussed in Biotechnology
11: 998-1000, 1117-9. Encouraging progress on trials of antibodies to reduce arthritis
is reported in F.H. Durie et al., "Prevention of collagen-induced arthritis with
an antibody to gp39, the ligand for CD40", Science
261: 1328-30, 1669-70. A book on the subject is Specific Immunotherapy of Cancer
with Vaccines, eds., J.-C. Bystryn et al., Annals of the NY Academy of Sciences
690: 1-402. A special issue of Scientific American
(Sept 1993) is on the immune system.
The modulation of a designed protein by metal ions is reported in Science 261: 879-85. The use of combinatorial mutagenesis to engineer multiple properties in an enzyme is reported in PNAS 90: 8367-71. The use of mammalian cell culture for producing recombinant proteins is reported in Biotechnology 11: 1037-41.
The search for RNA catalysts is reviewed in Nature 365: 204-5; and D.P. Bartel & J.W. Szostak, "Isolation of new ribozymes from a large pool of random sequences", Science 261: 1411-8, 1402-3.
Molecular motors are discussed in Nature 365: 203; Science 261: 1112-3. A physical method for construction of a synthetic biopolymer is reported in Science 261: 1303-5. Controlled delivery of drugs is discussed in GEN (1 Sept 1993), 1, 20-1.